Name: the mplex protocol for multi-omic analyses of soil samples
Uploaded: 2018-05-30T14:00:00
Description: Watch this Scientific Journal Video about The MPLEx Protocol for Multi-omic Analyses of Soil Samples at JoVE.com

Os autores gostaria de agradecer Nathan Johnson por sua assistência na elaboração de figuras. Esta pesquisa foi apoiada pelo programa Pan-omics que é financiado pelo departamento de energia dos Estados Unidos escritório de biológicos e pesquisa ambiental (programa de ciência genômica), o Microbiomes no desenvolvimento de pesquisas dirigido laboratório de transição (menta) Iniciativa no Pacific Northwest National Laboratory, bem como o nacional institutos de saúde nacionais Instituto de Ciências de saúde ambiental (R01 ES022190) e NIH (P42 ES027704). KEBJ gostaria de agradecer HD084788 R21 apoio financeiro desenvolver e validar as técnicas de extração multi-omic romance. Este trabalho foi realizado na W. R. Wiley ambiental Molecular Sciences Laboratory (EMSL), uma instalação de usuário científico nacional DOE no laboratório nacional Noroeste Pacífico (PNNL). PNNL é um programa multi laborátório operado pela Battelle para o DOE sob contrato DE-AC06-76RL01830.

Nota: Os solos muito molhados podem ser liofilizados antes da extracção sem detrimento para a eficácia da extração. Solo úmido também pode ser usado mas deve ser considerado ao adicionar os reagentes em proporções específicas.
Nota: É recomendável usar 20g de peso do solo seco por extração, que deve ser dividido entre dois tubos de 50 mL (máximo de solo de 10 g por tubo de 50 mL). As extrações podem ser escaladas cima ou para baixo depende do amostra disponível.
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<ol>
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P., 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=Metabolomic+response+of+human+skin+tissue+to+low+dose+ionizing+radiation.">Metabolomic response of human skin tissue to low dose ionizing radiation.</a> Mol Biosyst. 8, 1979-1986 (2012).</li><li>Perera, R., 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=Dengue+virus+infection+perturbs+lipid+homeostasis+in+infected+mosquito+cells.">Dengue virus infection perturbs lipid homeostasis in infected mosquito cells.</a> PLoS Pathog. 8, e1002584 (2012).</li><li>Gao, X., 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=A+reversed-phase+capillary+ultra-performance+liquid+chromatography-mass+spectrometry+(UPLC-MS)+method+for+comprehensive+top-down/bottom-up+lipid+profiling.">A reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling.</a> Anal Bioanal Chem. 402, 2923-2933 (2012).</li><li>Sorensen, C. 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L., 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=Systems+virology+identifies+a+mitochondrial+fatty+acid+oxidation+enzyme,+dodecenoyl+coenzyme+A+delta+isomerase,+required+for+hepatitis+C+virus+replication+and+likely+pathogenesis.">Systems virology identifies a mitochondrial fatty acid oxidation enzyme, dodecenoyl coenzyme A delta isomerase, required for hepatitis C virus replication and likely pathogenesis.</a> J Virol. 85, 11646-11654 (2011).</li><li>Manza, L. L., Stamer, S. L., Ham, A. J., Codreanu, S. G., Liebler, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sample+preparation+and+digestion+for+proteomic+analyses+using+spin+filters.">Sample preparation and digestion for proteomic analyses using spin filters.</a> Proteomics. 5, 1742-1745 (2005).</li><li>Wisniewski, J. R., Zougman, A., Nagaraj, N., Mann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Universal+sample+preparation+method+for+proteome+analysis.">Universal sample preparation method for proteome analysis.</a> Nat Methods. 6, 359-362 (2009).</li><li>Zhou, J. Y., 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=Simple+sodium+dodecyl+sulfate-assisted+sample+preparation+method+for+LC-MS-based+proteomics+applications.">Simple sodium dodecyl sulfate-assisted sample preparation method for LC-MS-based proteomics applications.</a> Anal Chem. 84, 2862-2867 (2012).</li><li>Anderson, J. 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É importante notar que nem todos os laboratórios terão o mesmo equipamento disponível para que certos métodos, por exemplo, a etapa de Lise, podem ser adaptados. Aqui usamos num Vortex e sonicating, no entanto, o uso de um batedor de grânulo grande 50ml funcionaria. Se não houver um Liofilizador com uma temperatura do coletor capaz de-105 º C, as amostras podem ser secos sob uma corrente de azoto. Também os tipos de solo variam muito e podem incluir areia, silte, argila, turfa e barro (etc), e também p...

Avaliar as comunidades microbianas do solo tem implicações importantes para a compreensão de carbono ciclismo e mudança climática. Estudos recentes no entanto evidenciaram dificuldades, como a falta de genomas sequenciadas para microbiota em vários tipos de solo e a função desconhecida de muitas das proteínas detectadas. Estes resultam de desafios devido ao solo, sendo a mais complexa comunidade microbiana conhecida até à data de1,2,3. Análises multi-omic, que combinam os resultados de estudos de lipidomic, metatranscriptomic, metaproteomic, metabolómica e metagenomic, recentemente têm sido implementadas em numerosos estudos de solo, para uma maior compreensão os micróbios presentes, enquanto obter informações completas sobre as alterações moleculares que ocorrem devido a perturbações ambientais1,4,5. Um desafio com multi-omic estudos que é a espectrometria de massa (MS)-medições com base de metaproteomic, lipidomic e metabolómica normalmente requerem um processo de extração específico para cada omic ser MS compatível6,7 , 8 , 9. estes procedimentos precisos fazem sua implementação extremamente difícil ou impossível quando somente uma quantidade limitada de amostra está disponível. Esses desafios têm nos levou a investigar uma simultânea metabólito, proteínas e lipídios (mplexas) método da extração capaz de usando pequenos volumes de amostra ou massas, melhorar a precisão e a realização de preparações de amostra mais rápidas para todos os três análises 10. até à data, não existem procedimentos alternativos solo extração que podem atingir todos esses objetivos.
Para habilitar o multi-omic global análises de uma amostra de solo único, um protocolo de extração com solvente orgânico baseado em clorofórmio, metanol e água separações foi utilizado10. Este método foi originalmente desenvolvido para extrações de lipídios totais9,11 e mais recentemente foi alterado para a extração simultânea de metabolitos, proteínas e lipídios de uma única amostra12,13 ,14,15,16,17,18,19,20,21,22, 23,24,25,26,,27,28,29,30, permitindo menos quantidade de amostra e variabilidade experimental10. No protocolo de mplexas, o clorofórmio não é miscível com a água, que fornece a base para a trifásico de separação química dos constituintes da amostra em fracções distintas. A fase aquosa superior, portanto, contém os metabolitos hidrofílicos, seguidos por um disco de proteínas e em seguida uma camada lipídica na fase de clorofórmio inferior (Figura 1). Quando mplexas é aplicada a maioria dos solos, detritos de partículas acumula-se na parte inferior dos tubos de amostragem e podem ser descartado após a coleta todas as camadas. Cada tipo de solo pode ser diferente, no entanto e em solos altamente orgânicos tais como turfa, os detritos do solo permanece na camada média e não cair no fundo do tubo de amostragem. Mplexas oferece várias vantagens ao isolar a molécula vários tipos da mesma amostra como 1) pequenas quantidades de amostra podem ser usadas para análises de multi-omic, 2) multi-omic extrações da mesma amostra diminuição geral experimental variabilidade, e 3) o maior número de amostras pode ser preparado muito mais rápido para maior taxa de transferência de estudos10. Juntos, esses benefícios são vitais para fornecer melhores recursos de medição para avaliar amostras de solo e suas comunidades microbianas complexas.

<table>
	<tbody>
		<tr>
			<td>Chloroform</td>
			<td>Sigma-Aldrich</td>
			<td>650498</td>
			<td>Stored at -20&#176;C !Caution chloroform has acute potential health effects, skin irritation and possible chemical burns, irritation to the respiratory system, may affect the kidneys, liver, heart. Wear suitable protective glasses, clothing and gloves, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Sigma-Aldrich</td>
			<td>34860</td>
			<td>Stored at -20&#176;C !Caution Methanol may cause respiratory tract, skin and eye irritation, may damage the nerves, kidneys and liver. Wear suitable protective glasses, clothing and gloves, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Purified water from Millipore</td>
			<td>Milli-Q</td>
			<td></td>
			<td>Water purification system.</td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulfate</td>
			<td>Sigma-Aldrich</td>
			<td>L6026</td>
			<td>!Caution SDS causes acute toxicity and is flammable. It is a skin, eye and airway irritant. Wear gloves and safety glasses.</td>
		</tr>
		<tr>
			<td>Soil protein extraction kit</td>
			<td>MoBio, NoviPure Soil Protein Extraction Kit, Qiagen</td>
			<td>30000-20</td>
			<td></td>
		</tr>
		<tr>
			<td>DL-dithiothreitol</td>
			<td>Sigma-Aldrich</td>
			<td>43815</td>
			<td></td>
		</tr>
		<tr>
			<td>1M Trizma HCL</td>
			<td>Sigma-Aldrich</td>
			<td>T2694</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloroacetic acid</td>
			<td>Sigma-Aldrich</td>
			<td>T0699</td>
			<td>!Caution TCA is caustic, toxic and may cause skin burns. Wear gloves and safety glasses.</td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>Sigma-Aldrich</td>
			<td>650501</td>
			<td>Stored at -20&#176;C !Caution Acetone may cause respiratory tract and skin and eye irritation. Flammable liquid and vapor. Wear safety glasses gloves and a lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Urea</td>
			<td>Sigma-Aldrich</td>
			<td>208884</td>
			<td>!Caution Urea is an eye and skin irritant, use gloves and safety glasses</td>
		</tr>
		<tr>
			<td>Ammonium bicarbonate</td>
			<td>Fluka</td>
			<td>09830</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin</td>
			<td>Promega</td>
			<td>V528A</td>
			<td>20&#181;g vials</td>
		</tr>
		<tr>
			<td>Bicinchoninic acid protein assay kit</td>
			<td>Pierce</td>
			<td>23227</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium Formate</td>
			<td>Sigma-Aldrich</td>
			<td>09735</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetonitrile</td>
			<td>Sigma-Aldrich</td>
			<td>34998</td>
			<td>!Caution Acetonitrile is a skin and eye irritant. Highly flammable. Wear gloves and safety glasses. Work in a fume hood.</td>
		</tr>
		<tr>
			<td>Trifluoroacetic acid</td>
			<td>Sigma-Aldrich</td>
			<td>T6508</td>
			<td>!Caution TFA is extremely hazardous in case of skin contact, eye contact, ingestion and inhalation. May produce tissue damage particularly on mucous membranes of eyes, mouth and respiratory tract. Skin contact may produce burns. Wear gloves, lab coat, safety glasses and work in a fume hood.</td>
		</tr>
		<tr>
			<td>Methoxyamine hydrochloride</td>
			<td>Sigma-Aldrich</td>
			<td>226904</td>
			<td>!Caution Methoxyamine hydrochloride causes severe burns and serious damage to eyes, may cause sensitization by skin contact. Wear safety glasses, gloves and lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Pyridine</td>
			<td>Sigma-Aldrich</td>
			<td>270970</td>
			<td>!Caution Pyridine can cause skin and eye irritation, central nervous system depression. Vapor may cause flash fire. Wear safety glasses, gloves and lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>N-Methyl-N-(trimethylsilyl)trifluoroacetamide with 1% trimethylchlorosilane</td>
			<td>Sigma-Aldrich</td>
			<td>69478</td>
			<td>!Caution MSTFA + 1% TMCS can cause skin corrosion, serious eye damage and specific target organ toxicity. Flammable liquid and vapor. Wear safety glasses, gloves and lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>P9541</td>
			<td></td>
		</tr>
		<tr>
			<td>Milli-Q water purification system</td>
			<td>Millipore</td>
			<td></td>
			<td>model MPGP04001</td>
		</tr>
		<tr>
			<td>Vortex</td>
			<td>Scientific Industries</td>
			<td>SI-0236</td>
			<td>Vortex Genie 2</td>
		</tr>
		<tr>
			<td>Probe sonicator</td>
			<td>FisherBrand</td>
			<td></td>
			<td>model FB505</td>
		</tr>
		<tr>
			<td>Refrigerated centrifuge</td>
			<td>Eppendorf</td>
			<td></td>
			<td>model 5810R</td>
		</tr>
		<tr>
			<td>50mL tube swinging bucket rotor</td>
			<td>Eppendorf</td>
			<td>A-4-44</td>
			<td></td>
		</tr>
		<tr>
			<td>50mL fixed angle rotor</td>
			<td>Eppendorf</td>
			<td>FA-45-6-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Balance</td>
			<td>OHAUS</td>
			<td></td>
			<td>model V22PWE150IT</td>
		</tr>
		<tr>
			<td>Serological pipette controller</td>
			<td>Eppendorf</td>
			<td>12-654-100</td>
			<td></td>
		</tr>
		<tr>
			<td>10mL, 25mL glass serological pipettes</td>
			<td>FisherBrand</td>
			<td>13-678-27F, 13-678-36D</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermomixer with Thermotop</td>
			<td>Eppendorf</td>
			<td>5382000015, 5308000003</td>
			<td></td>
		</tr>
		<tr>
			<td>0.9 &#8211; 2.0 mm blend stainless steel beads</td>
			<td>NextAdvance</td>
			<td>SSB14B</td>
			<td></td>
		</tr>
		<tr>
			<td>0.15 mm garnet beads</td>
			<td>MoBio</td>
			<td>13122-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic stir plate</td>
			<td>FisherBrand</td>
			<td>11-100-16SH</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic stir bar</td>
			<td>FisherBrand</td>
			<td>14512130</td>
			<td></td>
		</tr>
		<tr>
			<td>pH paper strips, pH range 0&#8211;14</td>
			<td>FisherBrand</td>
			<td>M95903</td>
			<td></td>
		</tr>
		<tr>
			<td>15mL, 50mL conical polypropylene centrifuge tube</td>
			<td>Genesee Scientific</td>
			<td>21-103 21-108</td>
			<td>chloroform compatible</td>
		</tr>
		<tr>
			<td>50mL vortex attachment</td>
			<td>MoBio</td>
			<td>13000-V1-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Ice bucket</td>
			<td>FisherBrand</td>
			<td>02-591-44</td>
			<td></td>
		</tr>
		<tr>
			<td>27.25x70mm glass vials</td>
			<td>FisherBrand</td>
			<td>03-339-22K</td>
			<td></td>
		</tr>
		<tr>
			<td>Breathe Easier plate membranes</td>
			<td>Midwest Scientific</td>
			<td>BERM-2000</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol wipes</td>
			<td>Diversified Biotech</td>
			<td>BPWP-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Heater shaker incubator</td>
			<td>Benchmark, Incu-Shaker Mini</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Analog rotisserie tube rotator</td>
			<td>SoCal BioMed, LLC</td>
			<td>82422001</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter-Aided-Sample-Prep kit</td>
			<td>FASP; Expedeon</td>
			<td>44250</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate reader</td>
			<td>Biotek, EPOCH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>-20 Degree Celsius Freezer</td>
			<td>Fisher</td>
			<td>13986149</td>
			<td></td>
		</tr>
		<tr>
			<td>-80 Degree Celsius Freezer</td>
			<td>Stirling Ultracold</td>
			<td>SU78OUE</td>
			<td></td>
		</tr>
		<tr>
			<td>Q-Exactive ion trap mass spectrometer</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Agilent 7890A gas chromatograph coupled with a single quadrupole 5975C mass spectrometer</td>
			<td>Agilent Technologies, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LTQ-Orbitrap Velo</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Waters NanoEquityTM UPLC system</td>
			<td>Millford, MA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>250mL media bottle</td>
			<td>FisherBrand</td>
			<td>1395-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Waters vial</td>
			<td>Waters</td>
			<td>186002805</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass MS sample vial and inserts</td>
			<td>MicroSolv</td>
			<td>9502S-WCV, 9502S-02ND</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass HPLC vial and snap caps</td>
			<td>MicroSolv</td>
			<td>9512C-0DCV, 9502C-10C-B</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC 96-well plate</td>
			<td>Agilent</td>
			<td>5042-6454</td>
			<td></td>
		</tr>
		<tr>
			<td>Large glass vial 27.25x70mm</td>
			<td>FisherBrand</td>
			<td>03-339-22K</td>
			<td></td>
		</tr>
		<tr>
			<td>Lyophilizer</td>
			<td>Labconco</td>
			<td>7934021</td>
			<td></td>
		</tr>
		<tr>
			<td>Polished stainless steel flat head spatula</td>
			<td>Spoonula; FisherBrand</td>
			<td>14-375-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Kim wipes</td>
			<td>Kimberly-Clark</td>
			<td>34721</td>
			<td></td>
		</tr>
		<tr>
			<td>XBridge C18, 250x4.6 mm, 5 &#956;M with 4.6x20 mm guard column</td>
			<td>Waters</td>
			<td>186003117, 186003064</td>
			<td></td>
		</tr>
		<tr>
			<td>Agilent 1100 series HPLC system</td>
			<td>Agilent Technologies</td>
			<td>G1380-90000</td>
			<td></td>
		</tr>
		<tr>
			<td>1.7mL centrifuge tube</td>
			<td>Sorenson</td>
			<td>11700</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton Glass Syringes, 5mL, 50&#181;L and 250&#181;L</td>
			<td>Hamilton</td>
			<td>81517, 80975, 81175</td>
			<td></td>
		</tr>
		<tr>
			<td>Pasteur Pipettes</td>
			<td>FisherBrand</td>
			<td>13-678-20A</td>
			<td></td>
		</tr>
		<tr>
			<td>Pasteur Pipette Bulbs</td>
			<td>Sigma-Aldrich</td>
			<td>Z111597</td>
			<td></td>
		</tr>
		<tr>
			<td>Bath Sonicator</td>
			<td>Branson 1800 Ultrasonic Cleaner</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum Centrifuge</td>
			<td colspan="2">Labconco Centrivap Acid-Resistant Concentrator System</td>
			<td></td>
		</tr>
		<tr>
			<td>MicroSpin Columns, C18 Silica</td>
			<td>The Nest Group</td>
			<td>SEM SS18V</td>
			<td></td>
		</tr>
	</tbody>
</table>

the mplex protocol for multi-omic analyses of soil samples

Espectrometria de massa (MS)-com base metaproteomic integrado, metabolómica e estudos lipidomic (multi-omic) estão transformando nossa capacidade de compreender e caracterizar as comunidades microbianas em sistemas ambientais e biológicos. Estas medições são mesmo permitindo análises reforçadas das comunidades microbianas complexo solo, que são os mais complexos sistemas microbianos conhecidos até à data. Multi-omic análises, no entanto, têm desafios de preparação de amostra, desde extrações separadas são normalmente necessários para cada estudo omic, amplificando assim consideravelmente o tempo de preparação e a quantidade de amostra necessária. Para resolver essa limitação, um método de 3-em-1 para a extração simultânea de metabolitos, proteínas e lipídios (mplexas) da mesma amostra de solo foi criado através da adaptação de uma abordagem baseada em solvente. Este protocolo de mplexas tem provado para ser simples e robusto para muitos tipos de amostra, mesmo quando utilizado para quantidades limitadas de amostras complexas. O método mplexas habilitado também extremamente rápida multi-omic medições necessárias para obter uma melhor compreensão dos membros de cada comunidade microbiana, ao avaliar as mudanças que ocorrem após perturbações biológicas e ambientais.

Quando o protocolo de mplexas foi usado para extrair as moléculas do solo de pradaria nativa de Kansas (um solo Mollisol), as análises de três vias previstas resultados 3376 peptídeos, 105 lipídios e 102 metabólitos polares (todas as identificações exclusivas). Enquanto o protocolo de mplexas tem sido bem estabelecido para geral extração de lipídios e metabolitos12,13,14,<su...

Watch this Scientific Journal Video about The MPLEx Protocol for Multi-omic Analyses of Soil Samples at JoVE.com

The MPLEx Protocol for Multi-omic Analyses of Soil Samples

Um protocolo é apresentado para simultaneamente extraindo metabolitos, proteínas e lipídios de uma amostra de solo único, permitindo tempos de preparação de amostra reduzida e multi-omic análises de espectrometria de massa das amostras com quantidades limitadas.

O protocolo de mplexas para Multi-omic análises de amostras de solo

Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and ...

Research

JoVE Journal

Chemistry

Transport of Surface-modified Carbon Nanotubes through a Soil Column

Transporte de nanotubos de carbono de superfície modificada através de uma coluna de solo

Carbon nanotubes (CNTs) are widely manufactured nanoparticles, which are being utilized in a number of consumer products, such as sporting goods, ...

Química

A Direct, Early Stage Guanidinylation Protocol for the Synthesis of Complex Aminoguanidine-containing Natural Products

Um direto, Early Stage Guanidinylation Protocolo para a Síntese de complexos contendo aminoguanidina Produtos Naturais

The guanidine functional group, displayed most prominently in the amino acid arginine, one of the fundamental building blocks of life, is an important ...

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Protocolo para a Síntese de Derivados de Anilina Orto-trifluorometoxilados

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of ...

A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating

Um protocolo para a produção de nanopartículas Gliadina-cianoacrilato para hidrofílico Coating

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray ...

A Simple and Efficient Protocol for the Catalytic Insertion Polymerization of Functional Norbornenes

Um protocolo simples e eficiente para a polimerização por inserção catalítica de norbornenos funcionais

Norbornene can be polymerized by a variety of mechanisms, including insertion polymerization whereby the double bond is polymerized and the bicyclic ...

A Protocol for Safe Lithiation Reactions Using Organolithium Reagents

Um protocolo para reações seguras litiação Usando organolï¿½io Reagentes

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents ...

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

Um protocolo otimizado para a radiomarcação eficiente de nanopartículas de ouro usando um<sup&gt; 125</sup&gt; Marcado com Eu Azida grupo prostético

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient ...

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles (PPAs) and Related Biomaterials

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles PPAs and Related Biomaterials

Facile protocolo para a síntese de auto montagem baseada em poliamina peptídeo Amphiphiles (CAE) e biomateriais relacionados

Polyamine-based Peptide Amphiphiles (PPAs) are a new class of self-assembling amphiphilic biomaterials-related to the peptide amphiphiles (PAs). ...

Automation of a Positron-emission Tomography (PET) Radiotracer Synthesis Protocol for Clinical Production

Automation of a Positron-emission Tomography PET Radiotracer Synthesis Protocol for Clinical Production

Automação de um protocolo de síntese Positron emission Tomography (PET) Radiotracer para produção clínica

The development of new positron-emission tomography (PET) tracers is enabling researchers and clinicians to image an increasingly wide array of biological ...

Biological Samples Preparation for Speciation at Cryogenic Temperature using High-Resolution  X-Ray Absorption Spectroscopy

Preparação de amostras biológicas para especiação em temperatura criogênica usando espectroscopia de absorção de raios-X de alta resolução

The study of elements with X-ray absorption spectroscopy (XAS) is of particular interest when studying the role of metals in biological systems. Sample ...