米国地質調査ハモンド湾生物ステーションの研究施設の利用、米魚類野生生物局と水産のスタッフと海カナダ海ヤツメウナギを提供するために感謝しますこの研究は、李維、柯李五大湖漁業委員会からの補助金によって支えられました。

<ol>
	<li>Wyatt, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pheromones and Animal Behavior: Chemical Signals and Signatures. , (2014).</li><li>Zhu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reverse+chemical+ecology:+Olfactory+proteins+from+the+giant+panda+and+their+interactions+with+putative+pheromones+and+bamboo+volatiles.">Reverse chemical ecology: Olfactory proteins from the giant panda and their interactions with putative pheromones and bamboo volatiles.</a> Proceedings of the National Academy of Sciences. 114 (46), E9802-E9810 (2017).</li><li>Leal, W. 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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Insect Pheromones and Their Use in Pest Management. , (2013).</li><li>Pizzolon, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=When+fathers+make+the+difference:+efficacy+of+male+sexually+selected+antimicrobial+glands+in+enhancing+fish+hatching+success.">When fathers make the difference: efficacy of male sexually selected antimicrobial glands in enhancing fish hatching success.</a> Functional Ecology. 24 (1), 141-148 (2010).</li><li>Stacey, N., Sorensen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Hormones in communication | Hormonal Pheromones Encyclopedia of Fish Physiology. , (2011).</li><li>Kobayashi, M., Sorensen, P. W., Stacey, N. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hormonal+and+pheromonal+control+of+spawning+behavior+in+the+goldfish.">Hormonal and pheromonal control of spawning behavior in the goldfish.</a> Fish Physiology and Biochemistry. 26 (1), 71-84 (2002).</li><li>Stacey, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hormonally-derived+pheromones+in+teleost+fishes.">Hormonally-derived pheromones in teleost fishes.</a> Fish Pheromones and Related Cues. , 33-88 (2015).</li><li>Kuhlisch, C., Pohnert, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolomics+in+chemical+ecology.">Metabolomics in chemical ecology.</a> Natural Product Reports. 32 (7), 937-955 (2015).</li><li>Prince, E. 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A., Bergstedt, R. A., Sorensen, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory-mediated+stream-finding+behavior+of+migratory+adult+sea+lamprey+(Petromyzon+marinus).">Olfactory-mediated stream-finding behavior of migratory adult sea lamprey (Petromyzon marinus).</a> Canadian Journal of Fisheries and Aquatic Sciences. 68, (2011).</li><li>Wagner, C. M., Jones, M. L., Twohey, M. B., Sorensen, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+field+test+verifies+that+pheromones+can+be+useful+for+sea+lamprey+(Petromyzon+marinus)+control+in+the+Great+Lakes.">A field test verifies that pheromones can be useful for sea lamprey (Petromyzon marinus) control in the Great Lakes.</a> Canadian Journal of Fisheries and Aquatic Sciences. 63 (3), 475-479 (2006).</li><li>Wagner, C. M., Twohey, M. B., Fine, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conspecific+cueing+in+the+sea+lamprey:+do+reproductive+migrations+consistently+follow+the+most+intense+larval+odour?.">Conspecific cueing in the sea lamprey: do reproductive migrations consistently follow the most intense larval odour?.</a> Animal Behaviour. 78, (2009).</li><li>Siefkes, M. J., Winterstein, S. R., Li, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+that+3-keto+petromyzonol+sulphate+specifically+attracts+ovulating+female+sea+lamprey+Petromyzon+marinus.">Evidence that 3-keto petromyzonol sulphate specifically attracts ovulating female sea lamprey Petromyzon marinus.</a> Animal Behaviour. 70, (2005).</li><li>Siefkes, M. J., Steeves, T. B., Sullivan, W. P., Twohey, M. 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Y., Schuhmacher, R., Krska, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liquid+chromatography-mass+spectrometry+for+the+determination+of+chemical+contaminants+in+food.">Liquid chromatography-mass spectrometry for the determination of chemical contaminants in food.</a> TRAC Trends in Analytical Chemistry. 59, 59-72 (2014).</li><li>Little, J. L., Williams, A. J., Pshenichnov, A., Tkachenko, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+&amp;#34;known+unknowns&amp;#34;+utilizing+accurate+mass+data+and+ChemSpider.">Identification of &amp;#34;known unknowns&amp;#34; utilizing accurate mass data and ChemSpider.</a> Journal of the American Society for Mass Spectrometry. 23 (1), 179-185 (2012).</li><li>Beckonert, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+profiling,+metabolomic+and+metabonomic+procedures+for+NMR+spectroscopy+of+urine,+plasma,+serum+and+tissue+extracts.">Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts.</a> Nature Protocols. 2 (11), 2692 (2007).</li><li>Kaiser, B., Wright, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Draft Bruker XRF Spectroscopy User Guide: Spectral Interpretation and Sources of Interference. , (2008).</li><li>Breitmaier, E., Sinnema, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Structure Elucidation by NMR in Organic Chemistry: A Practical Guide. , (1993).</li><li>Seco, J. 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J., Li, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discovery+and+characterization+of+natural+products+that+act+as+pheromones+in+fish.">Discovery and characterization of natural products that act as pheromones in fish.</a> Natural Product Reports. , (2018).</li><li>Yambe, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=L-Kynurenine,+an+amino+acid+identified+as+a+sex+pheromone+in+the+urine+of+ovulated+female+masu+salmon.">L-Kynurenine, an amino acid identified as a sex pheromone in the urine of ovulated female masu salmon.</a> Proceedings of the National Academy of Sciences. 103 (42), 15370-15374 (2006).</li><li>Rasmussen, L., Lee, T. D., Zhang, A., Roelofs, W. L., Daves, G. 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著者が明らかに何もありません。

魚は、化学の世界はまだ識別される化合物の完全に住んでいます。バイオアッセイ誘導分別識別しサクラマス31、アジア象32、および海ヤツメウナギ33で観察など、多くの化学的相互作用を仲介する生理活性分子を特徴付ける重要な証明されています 34,35。バイオアッセイ誘導分別が正確...

フェロモンは、食料源を検索、検出捕食者および個体1の社会的な相互作用を仲介することでそれらを支援する個人によってリリースされた特定の化学信号です。昆虫のフェロモン ・ コミュニケーションはずっとよく勉強2;しかし、化学的同定と水生脊椎動物フェロモンの生物学的機能が検討されていないとして広く。Id の知識とフェロモンのリリースの機能は、絶滅危惧種3,4または制御害虫種5,6の回復を容易に適用できます。これらの技術のアプリケーションは、分離・生理活性フェロモン成分の同定を必要とします。
フェロモンの同定は、天然物化学の枝です。フェロモン研究の進歩は、フェロモン分子自身の性質上、部分的に制限されています。フェロモンは、不安定になり、少量でリリースされた、揮発性7,8または9水溶性化合物の微量を検出するだけいくつかのサンプリング手法が存在します。フェロモン的なアプローチには、1) 知られている化合物、2) メタボロミクス、および 3) バイオアッセイ誘導分別の対象となるスクリーニングが含まれます。知られている化合物の標的スクリーニングは、フェロモンとして機能するいると仮定の生理的過程の市販の代謝の副産物をテストします。研究者のみ知られており、利用可能な化合物をテストできますので、この方法は制限されています。ただし、それをもたらしました金魚の性ホルモンの成功した識別関数フェロモン10、11,12として。メタボロミクスは、潜在的な生物学的システム13内低分子代謝産物を区別する第 2 フェロモン同定アプローチです。2 つのグループ (すなわちアクティブと非アクティブのエキス) の代謝プロファイルの比較により、潜在的な代謝プロファイルの id 構造の解明から代謝物を精製し、生物活性は確認された14です。特定の混合物の複雑な配合の添加剤または相乗効果、代謝物は一緒にではなく、分数13のシリーズとしてと見なされるために、メタボロミクスで検出される可能性が高い。まだ、メタボロミクスの実装は、結果のデータは、新規構造の解明を促進しないので合成参照の可用性に依存します。
バイオアッセイ誘導の分別、2 つのフィールドにまたがる統合、反復的なアプローチ: 化学および生物学。このアプローチは、分離とアクティブなフェロモン化合物の同定をガイドするのに生理・行動生物検定の結果を使用します。粗野なエキスは、 (すなわち、分子サイズ、極性等)の化学的性質によって分別され、生物検定および/または電気 olfactogram (EOG) 録音でのテストです。生理活性成分をスクリーニング、分別および EOGs および/または生物検定の手順を繰り返ししています。分子量と合成される化合物のテンプレートを生成する構造情報提供分光法と分光学的方法により純粋な活性化合物の構造を明らかにします。バイオアッセイ誘導の分別には、多様な代謝と生合成の経路から予測される可能性が高いではないユニークな化学のスケルトンと潜在的新規フェロモンをもたらすことができます。
ここでは、分離し、男性の海ヤツメウナギ セックス フェロモン化合物の生物活性を評価するために使用するバイオアッセイ誘導分別のプロトコルについて述べる。海ヤツメウナギ (カワヤツメ属 marinus) は、これらの魚は大きく 3 つの段階から成る、さくの生活史を仲介する化学手掛かりの嗅覚の検出に依存するため、フェロモン ・ コミュニケーションを研究する理想的な脊椎動物のモデルです。幼虫、少年、大人。海ヤツメウナギ淡水ストリームの底に穴を掘る、抜本的な変態、湖または海洋大ホスト魚を寄生彼らに移行する少年に変換します。ホストの魚からデタッチ後大人は産卵ストリームに戻って移行, ストリーム居住者幼虫15,16,17,18,19 発表渡り鳥のフェロモンに導かれて.成熟した雄は産卵場に登る、仲間を集めるし、約一週間、断続的に産卵して死ぬ15,20の多成分性フェロモンをリリースします。海ヤツメウナギ フェロモンの同定は、フェロモン通信システムの変調がラウレンツィアーナ五大湖21の侵襲の海ヤツメウナギにオプションの中では重要です。

<table border="0" cellpadding="0" cellspacing="0" width="1135">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:340px;">Premium standard wall borosilicate capillaries with filament&#160;</td>
			<td style="width:199px;">Warner Instruments</td>
			<td style="width:160px;">G150F-4</td>
			<td style="width:436px;">recording and reference electrode (OD 1.5 mm, ID 0.86 mm)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pipette puller instrument&#160;</td>
			<td style="width:199px;">Narishige</td>
			<td style="width:160px;">PC-10</td>
			<td>pulls electrodes for EOGs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Diamond-tipped glass cutter</td>
			<td style="width:199px;">Generic</td>
			<td style="width:160px;"></td>
			<td>cut tip of electrodes for EOG</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Borosilicate glass capillaries</td>
			<td style="width:199px;">World Precision Instruments</td>
			<td style="width:160px;">1B150-4</td>
			<td>odorant delivery tube for EOG</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:340px;">Recording electrode holder E Series straight body with Ag/AgCl pellet for glass capillary OD 1.5 mm</td>
			<td style="width:199px;">Warner Instruments</td>
			<td style="width:160px;">ESP-M15N</td>
			<td>recording electrode holder</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:340px;">Reference electrode holder E Series with handle with&#160; Ag/AgCl pellet&#160; for glass capillary OD 1.5 mm</td>
			<td style="width:199px;">Warner Instruments</td>
			<td style="width:160px;">E45P-F15NH</td>
			<td>reference electrode holder</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">1 mm pin</td>
			<td style="width:199px;">Warner Instruments</td>
			<td style="width:160px;">WC1-10</td>
			<td>to bridge reference and recording electrode holders</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">2 mm pin</td>
			<td style="width:199px;">Warner Instruments</td>
			<td style="width:160px;">WC2-5</td>
			<td>to bridge reference and recording electrode holders</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Agar</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">A1296</td>
			<td>molten agar to fill electrodes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Potassium chloride (KCl)</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">P9333</td>
			<td>3M KCl to fill electrodes and electrode holders</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Micropipette microfil</td>
			<td style="width:199px;">World Precision Instruments</td>
			<td style="width:160px;">MF28G-5</td>
			<td>to fill electrodes and electrode holders&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">L-Arginine</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">A5006</td>
			<td>positive control odorant for EOG</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Methanol</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">34860</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Water bath</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td>holds odorants for EOG</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3-aminobenzoic acid ethyl ester (MS222)</td>
			<td style="width:199px;">Syndel USA</td>
			<td>Tricaine1G</td>
			<td>EOG anesthetic&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Gallamine triethiodide</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">G8134-5G</td>
			<td>EOG paralytic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">1 mL syringe</td>
			<td style="width:199px;">BD Biosciences</td>
			<td>301025</td>
			<td>to administer paralytic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Subcutaneous needle 26G 5/8</td>
			<td style="width:199px;">BD Biosciences</td>
			<td style="width:160px;">305115</td>
			<td>to administer paralytic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Roller clamp</td>
			<td style="width:199px;">World Precision Instruments</td>
			<td style="width:160px;">14043-20</td>
			<td>adjust flow rate of anesthic into lamprey&#39;s mouth</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Sodium chloride (NaCl)</td>
			<td style="width:199px;">J.T. Baker</td>
			<td style="width:160px;">3624-05</td>
			<td>for preparation of 0.9% saline</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">V-shaped plastic stand as specimen stage</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td>holds lamprey during EOG</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Plastic trough</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td>holds V-shaped plastic stand during EOG</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Scalpel Blades - #11</td>
			<td style="width:199px;">Fine Science Tools</td>
			<td style="width:160px;">10011-00</td>
			<td>for EOG dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Scalpel Handle - #3</td>
			<td style="width:199px;">Fine Science Tools</td>
			<td style="width:160px;">10003-12</td>
			<td>for EOG dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Straight ultra fine forceps</td>
			<td style="width:199px;">Fine Science Tools</td>
			<td style="width:160px;">11252-00</td>
			<td>for EOG dissection, Dumont #5SF Forceps</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Curved ultra fine forceps</td>
			<td style="width:199px;">Fine Science Tools</td>
			<td>11370-42</td>
			<td>for EOG dissection, Moria MC40B</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Straight pring Scissors</td>
			<td style="width:199px;">Fine Science Tools</td>
			<td>15003-08</td>
			<td>for EOG dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Stereomicroscope</td>
			<td style="width:199px;">Zeiss</td>
			<td style="width:160px;">Discovery V8</td>
			<td>for EOG dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Illuminator light</td>
			<td style="width:199px;">Zeiss</td>
			<td style="width:160px;">CL 1500 ECO</td>
			<td>for EOG dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Plastic tubing</td>
			<td>Generic</td>
			<td style="width:160px;"></td>
			<td>to connect re-circulating EOG setup and water baths</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Odorant delivery tubing&#160;</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">In line filter and gasket set</td>
			<td style="width:199px;">Lee Company</td>
			<td style="width:160px;">TCFA1201035A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Micromanipulators</td>
			<td style="width:199px;">Narishige</td>
			<td style="width:160px;">MM-3</td>
			<td>to position electrodes and odorant delivery capillary tube</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Magnetic holding devices</td>
			<td style="width:199px;">Kanetec</td>
			<td style="width:160px;">MB-K</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Valve driver</td>
			<td style="width:199px;">Arduino</td>
			<td style="width:160px;">custom made</td>
			<td>to control the opening of the valve for odor stimulation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Electromagnetic valve</td>
			<td style="width:199px;">Lee Company</td>
			<td style="width:160px;">LFAA1201618H</td>
			<td>valve for odor stimulation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">NeuroLog AC/DC amplifier</td>
			<td>Digitimer Ltd.</td>
			<td style="width:160px;">NL106</td>
			<td>to increase the amplitude of the elictrical signal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">NeuroLog DC pre-amplifier with headstage</td>
			<td>Digitimer Ltd.</td>
			<td>NL102G</td>
			<td>to increase the amplitude of the elictrical signal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Low-pass 60 Hz filter</td>
			<td>Digitimer Ltd.</td>
			<td>NL125</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:340px;">Digitizer</td>
			<td>Molecular Devices LLC</td>
			<td style="width:160px;">Axon Digidata 1440A</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:340px;">Dell computer (OptiPlex 745) running Axoscope data acquistion software</td>
			<td style="width:199px;">Molecular Devices LLC</td>
			<td>AxoScope version 10.4&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Faraday cage</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td>Electromagnetic noise shielding</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Two-choice maze</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td>waterproofed marine grade plywood covered with plastic liner</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Trash pump</td>
			<td style="width:199px;">Honda</td>
			<td>WT30XK4A</td>
			<td>fills maze with water from nearby river</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:340px;">Peristaltic pump with tubing</td>
			<td style="width:199px;">Cole Parmer</td>
			<td style="width:160px;">Masterflex 07557-00</td>
			<td>to adminster odorants in maze</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Inverter Generator&#160;</td>
			<td style="width:199px;">Honda</td>
			<td style="width:160px;">EU1000i</td>
			<td>powers perstaltic pump</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Release cage</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:160px;">N/A</td>
			<td>used to acclimate lamprey in the maze</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Mesh</td>
			<td>Generic</td>
			<td style="width:160px;"></td>
			<td>used to contain the dimensions of the maze and minimize water turbulance with mesh rollers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Buckets (5 gallon)</td>
			<td>Generic</td>
			<td style="width:160px;"></td>
			<td>to mix odorants</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Flow meter</td>
			<td style="width:199px;">Marsh-McBirney</td>
			<td>Flo-Mate 2000</td>
			<td>to measure discharge</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">XAD 7 HP resin</td>
			<td style="width:199px;">Dow chemical</td>
			<td style="width:160px;">37380-43-1</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Methanol</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">34860</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Water bath</td>
			<td style="width:199px;">Yamato</td>
			<td style="width:160px;">BM 200</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:340px;">Freeze dryer</td>
			<td style="width:199px;">Labconco</td>
			<td style="width:160px;">CentriVap&#160; Concentrator</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">chloroform</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">CX1050</td>
			<td>for isolation of fraction pools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Silica gel 70-230 mesh</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">112926-00-8</td>
			<td>for isolation of fraction pools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Silica gel 230-400 mesh</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">112926-00-8</td>
			<td>for isolation of fraction pools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Pre-coated silica gel TLC plates</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">99571</td>
			<td>for isolation of fraction pools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">anisaldehyde</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">A88107</td>
			<td>for isolation of fraction pools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sephadex LH-20</td>
			<td style="width:199px;">GE Healthcare</td>
			<td style="width:160px;">17-0090-01</td>
			<td>for isolation of fraction pools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Amberlite XAD 7 HP resin</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">XAD7HP</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4, 2.5L capacity glass columns</td>
			<td>Ace Glass Inc.</td>
			<td>5820</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Acetone</td>
			<td style="width:199px;">Sigma</td>
			<td style="width:160px;">650501</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TQ-S TOF LC Mass spectrometer (or equivalent)</td>
			<td style="width:199px;">Waters Co.</td>
			<td style="width:160px;">N/A</td>
			<td>for structure elucidation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Binary HPLC pump</td>
			<td style="width:199px;">Waters Co.</td>
			<td style="width:160px;">1525</td>
			<td>for isolation of fraction pools/compounds</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Agilent NMR spectrometer, 900MHz (or equivalent)</td>
			<td style="width:199px;">Agilent</td>
			<td style="width:160px;">N/A</td>
			<td>for structure elucidation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rotovap drying system</td>
			<td style="width:199px;">Buchi</td>
			<td style="width:160px;">RII</td>
			<td>for extraction of conditioned water&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UV lamp (254 nm)</td>
			<td style="width:199px;">Spectronics Co.</td>
			<td style="width:160px;">ENF-240C</td>
			<td>for thin layer chromatography&#160;</td>
		</tr>
	</tbody>
</table>

the identification of sea lamprey pheromones using bioassay-guided fractionation

バイオアッセイ誘導の分別は、生理・行動生物検定の結果を使用して分離とアクティブなフェロモン化合物の同定をガイドする反復的なアプローチです。このメソッドは、幅広い動物種のフェロモンとしてその関数化学信号の正常性状に起因しました。海ヤツメウナギは行動または生理学的な応答を仲介するフェロモンを検出する嗅覚に頼る。私たちはこの魚の生物学の知識を使用して推定されるフェロモンの機能を仮定するし分離とアクティブなフェロモン成分の同定。クロマトグラフィーは、抽出、集中、およびエアコンの水から化合物の分離に使用されます。電気 olfactogram (EOG) 録音する分数は嗅覚応答を引き出すを決定するため行われています。2 つの選択肢迷路行動アッセイは、臭気の分数のいずれかがアクティブな行動もし、好みを誘発するかどうかを決定する使用されます。分光法と分光学的方法は、分子量や構造解明を支援する構造の情報を提供します。純粋な化合物の生物活性を EOG と行動実験で確認しました。迷路の中にみられる行動応答は最終的に自然のストリーム設定の機能を確認するためのフィールド設定で検証しなければなりません。これらの生物検定は、1) 分別プロセスをガイドし、2) 確認するデュアル役割と、分離されたコンポーネントの活性の定義します。ここでは、分別のバイオアッセイ誘導アプローチの有用性を例証する海ヤツメウナギ フェロモン同定の代表的な結果を報告します。海ヤツメウナギ フェロモンの同定は、フェロモン通信システムの変調が侵襲的海ヤツメウナギ ラウレンツィアーナ五大湖にオプションの中では特に重要です。このメソッドは、イチイの広範な配列の化学交信を特徴付けるし、水系の化学生態学に光を当てるに容易に適応できます。

バイオアッセイ誘導分別のプロトコルで説明されている手順をまとめた図を図 1に示します。プロトコルには、分離し、構造、嗅覚の効力 5 推定海ヤツメウナギ フェロモン (図 2) の行動の活性を評価する手順が含まれます。質量分析および NMR データ (図 3および図 4) を使用して...

Watch this Scientific Journal Video about The Identification of Sea Lamprey Pheromones Using Bioassay-Guided Fractionation at JoVE.com

The Identification of Sea Lamprey Pheromones Using Bioassay-Guided Fractionation

ここでは、分離し、構造、嗅力海ヤツメウナギの推定フェロモン化合物の行動応答を評価するためのプロトコルを提案する.

バイオアッセイ誘導分別を使用して海ヤツメウナギ フェロモンの同定

Bioassay-guided fractionation is an iterative approach that uses the results of physiological and behavioral bioassays to guide the isolation and ...

the-identification-sea-lamprey-pheromones-using-bioassay-guided

Research

JoVE Journal

Biochemistry

8.9K ビュー.  Michigan State University. ここでは、分離し、構造、嗅力海ヤツメウナギの推定フェロモン化合物の行動応答を評価するためのプロトコルを提案する.

ビデオ: The Identification of Sea Lamprey Pheromones Using Bioassay-Guided Fractionation

Identification of Fatty Acids in Bacillus cereus

The Bacillus species contain branched chain and unsaturated fatty acids (FAs) with diverse positions of the methyl branch (iso or anteiso) and of the double bond. Changes in FA composition play a crucial role in the adaptation of bacteria to their environment. These modifications entail a change in the ratio of iso versus anteiso branched FAs, and in the proportion of unsaturated FAs relative to saturated FAs, with double bonds created at specific positions. Precise identification of the FA profile is necessary to understand the adaptation mechanisms of Bacillus species.
Many of the FAs from Bacillus are not commercially available. The strategy proposed herein identifies FAs by combining information on the retention time (by calculation of the equivalent chain length (ECL)) with the mass spectra of three types of FA derivatives: fatty acid methyl esters (FAMEs), 4,4-dimethyl oxazoline derivatives (DMOX), and 3-pyridylcarbinyl ester (picolinyl). This method can identify the FAs without the need to purify the unknown FAs.
Comparing chromatographic profiles of FAME prepared from Bacillus cereus with a commercial mixture of standards allows for the identification of straight-chain saturated FAs, the calculation of the ECL, and hypotheses on the identity of the other FAs. FAMEs of branched saturated FAs, iso or anteiso, display a constant negative shift in the ECL, compared to linear saturated FAs with the same number of carbons. FAMEs of unsaturated FAs can be detected by the mass of their molecular ions, and result in a positive shift in the ECL compared to the corresponding saturated FAs.
The branching position of FAs and the double bond position of unsaturated FAs can be identified by the electron ionization mass spectra of picolinyl and DMOX derivatives, respectively. This approach identifies all the unknown saturated branched FAs, unsaturated straight-chain FAs and unsaturated branched FAs from the B. cereus extract.

Identification of Fatty Acids in Bacillus cereus

We propose a protocol to identify fatty acids without the need to purify them. It combines information on the retention times with the mass spectra of three types of fatty acid derivatives: fatty acid methyl esters (FAMEs), 4,4-dimethyl oxazoline derivatives (DMOX), and 3-pyridylcarbinyl esters (picolinyl).

脂肪酸の同定で<em&gt;セレウス菌</em

The Bacillus species contain branched chain and unsaturated fatty acids (FAs) with diverse positions of the methyl branch (iso or anteiso) and of the ...

生化学

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Aggregates of the neuronal Tau protein are found inside neurons of Alzheimer's disease patients. Development of the disease is accompanied by increased, abnormal phosphorylation of Tau. In the course of the molecular investigation of Tau functions and dysfunctions in the disease, nuclear magnetic resonance (NMR) spectroscopy is used to identify the multiple phosphorylations of Tau. We present here detailed protocols of recombinant production of Tau in bacteria, with isotopic enrichment for NMR studies. Purification steps that take advantage of Tau's heat stability and high isoelectric point are described. The protocol for in vitro phosphorylation of Tau by recombinant activated ERK2 allows for generating multiple phosphorylations. The protein sample is ready for data acquisition at the issue of these steps. The parameter setup to start recording on the spectrometer is considered next. Finally, the strategy to identify phosphorylation sites of modified Tau, based on NMR data, is explained. The benefit of this methodology compared to other techniques used to identify phosphorylation sites, such as immuno-detection or mass spectrometry (MS), is discussed.

We describe here a method to identify multiple phosphorylations of an intrinsically disordered protein by Nuclear Magnetic Resonance Spectroscopy (NMR), using Tau protein as a case study. Recombinant Tau is isotopically enriched and modified in vitro by a kinase prior to data acquisition and analysis.

本質的に無秩序なタンパク質の複数のリン酸化の同定のための核磁気共鳴分光法

Aggregates of the neuronal Tau protein are found inside neurons of Alzheimer's disease patients. Development of the disease is accompanied by increased, ...

A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis

Transcriptional coregulators are vital to the efficient transcriptional regulation of nuclear chromatin structure. Coregulators play a variety of roles in regulating transcription. These include the direct interaction with transcription factors, the covalent modification of histones and other proteins, and the occasional chromatin conformation alteration. Accordingly, establishing relatively quick methods for identifying proteins that interact within this network is crucial to enhancing our understanding of the underlying regulatory mechanisms. LC-MS/MS-mediated protein binding partner identification is a validated technique used to analyze protein-protein interactions. By immunoprecipitating a previously-identified member of a protein complex with an antibody (occasionally with an antibody for a tagged protein), it is possible to identify its unknown protein interactions via mass spectrometry analysis. Here, we present a method of protein preparation for the LC-MS/MS-mediated high-throughput identification of protein interactions involving nuclear cofactors and their binding partners. This method allows for a better understanding of the transcriptional regulatory mechanisms of the targeted nuclear factors.

We have established a method for the purification of coregulatory interaction proteins using the LC-MS/MS system.

LC-MS / MS分析を用いた核補因子と相互作用するタンパク質のハイスループット同定のためのタンパク質調製方法

Transcriptional coregulators are vital to the efficient transcriptional regulation of nuclear chromatin structure. Coregulators play a variety of roles in ...

Fractionation for Resolution of Soluble and Insoluble Huntingtin Species

The accumulation of misfolded proteins is central to pathology in Huntington's disease (HD) and many other neurodegenerative disorders. Specifically, a key pathological feature of HD is the aberrant accumulation of mutant HTT (mHTT) protein into high molecular weight complexes and intracellular inclusion bodies composed of fragments and other proteins. Conventional methods to measure and understand the contributions of various forms of mHTT-containing aggregates include fluorescence microscopy, western blot analysis, and filter trap assays. 
However, most of these methods are conformation specific, and therefore may not resolve the full state of mHTT protein flux due to the complex nature of aggregate solubility and resolution.
For the identification of aggregated mHTT and various modified forms and complexes, separation and solubilization of the cellular aggregates and fragments is mandatory. Here we describe a method to isolate and visualize soluble mHTT, monomers, oligomers, fragments, and an insoluble high molecular weight (HMW) accumulated mHTT species. HMW mHTT tracks with disease progression, corresponds with mouse behavior readouts, and has been beneficially modulated by certain therapeutic interventions1. This approach can be used with mouse brain, peripheral tissues, and cell culture but may be adapted to other model systems or disease contexts.

A method is described for fractionation of insoluble and soluble mutant huntingtin species from mouse brain and cell culture. The method described is useful for characterization and quantification of huntingtin protein flux and aids in analyzing protein homeostasis in disease pathogenesis and in the presence of perturbations

水溶性と不溶性の Huntingtin 種の解像度の分別

The accumulation of misfolded proteins is central to pathology in Huntington's disease (HD) and many other neurodegenerative disorders. Specifically, a ...

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and  GTPase-linked Immunosorbent Assay

The Rho GTPase family belongs to the Ras superfamily and includes approximately 20 members in humans. Rho GTPases are important in the regulation of diverse cellular functions, including cytoskeletal dynamics, cell motility, cell polarity, axonal guidance, vesicular trafficking, and cell cycle control. Changes in Rho GTPase signaling play an essential regulatory role in many pathological conditions, such as&#160;cancer, central nervous system diseases, and immune system-dependent diseases. The posttranslational modification of Rho GTPases (i.e., prenylation by mevalonate pathway intermediates) and GTP binding are key factors which affect the activation of this protein. In this paper, two essential and simple methods are provided to detect a broad range of Rho GTPase prenylation and GTP binding activities. Details of the technical procedures that have been used are explained step by step in this manuscript.

Here we describe a protocol to investigate the prenylation and guanosine-5'-triphosphate (GTP)-loading of Rho GTPase. This protocol consists of two detailed methods, namely membrane fractionation and a GTPase-linked immunosorbent assay. The protocol can be used for measuring the prenylation and GTP loading of different other small GTPases.

小さい gtp アーゼ プレニル化および分子量 GTP 結合を使用して膜分画と gtp アーゼ抗体法の検出

The Rho GTPase family belongs to the Ras superfamily and includes approximately 20 members in humans. Rho GTPases are important in the regulation of ...

Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

In this protocol we detail a method to obtain subcellular fractions of U937 cells without the use of ultracentrifugation or indiscriminate detergents. This method utilizes hypotonic buffers, digitonin, mechanical lysis and differential centrifugation to isolate the cytoplasm, mitochondria and plasma membrane. The process can be scaled to accommodate the needs of researchers, is inexpensive and straightforward. This method will allow researchers to determine protein localization in cells without specialized centrifuges and without the use of commercial kits, both of which can be prohibitively expensive. We have successfully used this method to separate cytosolic, plasma membrane and mitochondrial proteins in the human monocyte cell line U937.

Here, we present a protocol to isolate the plasma membrane, cytoplasm and mitochondria of U937 cells without the use of high-speed centrifugation. This technique can be used to purify subcellular fractions for subsequent examination of protein localization via immunoblotting.

高速遠心分離のない状態で U937 細胞のセル分別

In this protocol we detail a method to obtain subcellular fractions of U937 cells without the use of ultracentrifugation or indiscriminate detergents. ...

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

The study of kinase-substrate relationships is essential to gain a complete understanding of the functions of these enzymes and their downstream targets in both physiological and pathological states. CK2 is an evolutionarily conserved serine/threonine kinase with a growing list of hundreds of substrates involved in multiple cellular processes. Due to its pleiotropic properties, identifying and characterizing a comprehensive set of CK2 substrates has been particularly challenging and remains a hurdle in the study of this important enzyme. To address this challenge, we have devised a versatile experimental strategy that enables the targeted enrichment and identification of putative CK2 substrates. This protocol takes advantage of the unique dual co-substrate specificity of CK2 allowing for specific thiophosphorylation of its substrates in a cell or tissue lysate. These substrate proteins are subsequently alkylated, immunoprecipitated, and identified by liquid chromatography/tandem mass spectrometry (LC-MS/MS). We have previously used this approach to successfully identify CK2 substrates from Drosophila ovaries and here we extend the application of this protocol to human glioblastoma cells, illustrating the adaptability of this method to investigate the biological roles of this kinase in various model organisms and experimental systems.

The objective of this protocol is to label, enrich, and identify substrates of protein kinase CK2 from a complex biological sample such as a cell lysate or tissue homogenate. This method leverages unique aspects of CK2 biology for this purpose.

多彩な生化学的アプローチを用いた新規 CK2 キナーゼ基板の識別

The study of kinase-substrate relationships is essential to gain a complete understanding of the functions of these enzymes and their downstream targets ...

Identification of Orexin and Endocannabinoid Receptors in Adult Zebrafish Using Immunoperoxidase and Immunofluorescence Methods

Immunohistochemistry (IHC) is a highly sensitive and specific technique involved in the detection of target antigens in tissue sections with labeled antibodies. It is a multistep process in which the optimization of each step is crucial to obtain the optimum specific signal. Through IHC, the distribution and localization of specific biomarkers can be detected, revealing information on evolutionary conservation. Moreover, IHC allows for the understanding of expression and distribution changes of biomarkers in pathological conditions, such as obesity. IHC, mainly the immunofluorescence technique, can be used in adult zebrafish to detect the organization and distribution of phylogenetically conserved molecules, but a standard IHC protocol is not estasblished. Orexin and endocannabinoid are two highly conserved systems involved in the control of food intake and obesity pathology. Reported here are protocols used to obtain information about orexin peptide (OXA), orexin receptor (OX-2R), and cannabinoid receptor (CB1R) localization and distribution in the gut and brain of normal and diet-induced obese (DIO) adult zebrafish models. Also described are methods for immunoperoxidase and double immunofluorescence, as well as preparation of reagents, fixation, paraffin-embedding, and cryoprotection of zebrafish tissue and preparation for an endogenous activity-blocking step and background counterstaining. The complete set of parameters is obtained from previous IHC experiments, through which we have shown how immunofluorescence can help with the understanding of OXs, OX-2R, and CB1R distribution, localization, and conservation of expression in adult zebrafish tissues. The resulting images with highly specific signal intensity led to the confirmation that zebrafish are suitable animal models for immunohistochemical studies of distribution, localization, and evolutionary conservation of specific biomarkers in physiological and pathological conditions. The protocols presented here are recommended for IHC experiments in adult zebrafish.

Presented here are protocols for immunohistochemical characterization and localization of orexin peptide, orexin receptors, and endocannabinoid receptors in the gut and brains of normal and diet-induced obesity (DIO) adult zebrafish models using immunoperixidase and double immunofluorescence methods.

免疫ペルオキシダーゼおよび免疫蛍光法を用いて成体ゼブラフィッシュにおけるオレキシンおよびエンドカンナビノイド受容体の同定

Immunohistochemistry (IHC) is a highly sensitive and specific technique involved in the detection of target antigens in tissue sections with labeled ...

Measuring Interactions between Fluorescent Probes and Lignin in Plant Sections by sFLIM Based on Native Autofluorescence

In lignocellulosic biomass (LB), the activity of enzymes is limited by the appearance of non-specific interactions with lignin during the hydrolysis process, which maintains enzymes far from their substrate. Characterization of these complex interactions is thus a challenge in complex substrates such as LB. The method here measures molecular interactions between fluorophore-tagged molecules and native autofluorescent lignin, to be revealed by F&#246;rster resonance energy transfer (FRET). Contrary to FRET measurements in living cells using two exogenous fluorophores, FRET measurements in plants using lignin is not trivial due to its complex autofluorescence. We have developed an original acquisition and analysis pipeline with correlated observation of two complementary properties of fluorescence: fluorescence emission and lifetime. sFLIM (spectral and fluorescent lifetime imaging microscopy) provides the quantification of these interactions with high sensitivity, revealing different interaction levels between biomolecules and lignin.

This protocol describes an original setup combining spectral and fluorescence lifetime measurements to evaluate F&#246;rster resonance energy transfer (FRET) between rhodamine-based fluorescent probes and lignin polymer directly in thick plant sections.

ネイティブ自己蛍光に基づくsFLIMによる植物切片における蛍光プローブとリグニン間の相互作用の測定

In lignocellulosic biomass (LB), the activity of enzymes is limited by the appearance of non-specific interactions with lignin during the hydrolysis ...

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

This protocol describes a method to obtain subcellular protein fractions from mammalian cells using a combination of detergents, mechanical lysis, and isopycnic density gradient centrifugation. The major advantage of this procedure is that it does not rely on the sole use of solubilizing detergents to obtain subcellular fractions. This makes it possible to separate the plasma membrane from other membrane-bound organelles of the cell. This procedure will facilitate the determination of protein localization in cells with a reproducible, scalable, and selective method. This method has been successfully used to isolate cytosolic, nuclear, mitochondrial, and plasma membrane proteins from the human monocyte cell line, U937. Although optimized for this cell line, this procedure may serve as a suitable starting point for the subcellular fractionation of other cell lines. Potential pitfalls of the procedure and how to avoid them are discussed as are alterations that may need to be considered for other cell lines.

This fractionation protocol will allow researchers to isolate cytoplasmic, nuclear, mitochondrial, and membrane proteins from mammalian cells. The latter two subcellular fractions are further purified via isopycnic density gradient.

等密度勾配精製によるU937細胞の細胞分画

This protocol describes a method to obtain subcellular protein fractions from mammalian cells using a combination of detergents, mechanical lysis, and ...

バイオアッセイ誘導分別を使用して海ヤツメウナギ フェロモンの同定

バイオアッセイ誘導分別を使用して海ヤツメウナギフェロモンの同定