Name: linearization of the bradford protein assay
Uploaded: 2010-04-12T17:00:00
Description: Watch this Scientific Journal Video about Linearization of the Bradford Protein Assay at JoVE.com

本文是献给已故的博士兹维Selinger，谁主办的原创性研究，这里描述的内存。

Bradford蛋白校正曲线的线性化： 考马斯亮蓝蛋白测定，俗称Bradford法1，被广泛使用，因为其快速，便捷的协议，以及其相对灵敏度。不幸的是，有很大程度的曲率广泛的蛋白浓度（图1）。因此，只有一个比较高的蛋白浓度2-10 mg / ml的BSA，的狭窄的范围是用于校准曲线，更好地符合线性回归（图1，绿色）。然而，非线性需要未知样品的蛋白质浓度下降校准曲线的有限范...

<ol>
	<li>Bradford, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rapid+and+sensitive+method+for+the+quantitation+of+microgram+quantities+of+protein+utilizing+the+principle+of+protein-dye+binding.">A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.</a> Anal Biochem. 72, 248-254 (1976).</li><li>Chial, H. J., Thompson, H. B., Splittgerber, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+spectral+study+of+the+charge+forms+of+Coomassie+blue+G.">A spectral study of the charge forms of Coomassie blue G.</a> Anal Biochem. 209, 258-266 (1993).</li><li>Chial, H. J., Splittgerber, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+the+binding+of+Coomassie+brilliant+blue+to+proteins+at+low+and+neutral+pH.">A comparison of the binding of Coomassie brilliant blue to proteins at low and neutral pH.</a> Anal Biochem. 213, 362-369 (1993).</li><li>Compton, S. J., Jones, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+dye+response+and+interference+in+the+Bradford+protein+assay.">Mechanism of dye response and interference in the Bradford protein assay.</a> Anal Biochem. 151, 369-374 (1985).</li><li>Congdon, R. W., Muth, G. W., Splittgerber, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+binding+interaction+of+Coomassie+blue+with+proteins.">The binding interaction of Coomassie blue with proteins.</a> Anal Biochem. 213, 407-413 (1993).</li><li>Zor, T., Selinger, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Linearization+of+the+Bradford+protein+assay+increases+its+sensitivity:+theoretical+and+experimental+studies.">Linearization of the Bradford protein assay increases its sensitivity: theoretical and experimental studies.</a> Anal Biochem. 236, 302-308 (1996).</li></ol>

Bradford蛋白测定是受欢迎的，由于其性能和相对灵敏度缓解。在整个范围内通过协议获得的蛋白质浓度的线性化提出了进一步简化检测，作为未知样品不需要校准曲线的范围内下降。 更重要的是，进一步改进协议在原布拉德福德协议提供以下优点： <ol><li>提高准确度。 </li><li>灵敏度提高约一个量级，从而有可能确定在微孔板检测的牛血清白蛋白 6低至50纳克。 </li>...

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<td>Bradford reagent</td>
<td>&nbsp;</td>
<td>Bio-Rad Laboratories</td>
<td>500-0006</td>
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<td>Bovine Serum Albumin</td>
<td>&nbsp;</td>
<td>Amersco</td>
<td>0332</td>
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<td>Multiplate absorbance reader</td>
<td>&nbsp;</td>
<td>BioTek</td>
<td>Synergy2</td>
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linearization of the bradford protein assay

微克数量在布拉德福德考马斯亮蓝测定蛋白质的测定是通过在590 nm处测定吸光度。最常见的检测，使蛋白质在细胞裂解液，细胞组分或重组蛋白样品的快速和简单的量化，生化测定正常化的目的。然而，一种内在的非线性妥协这种方法的灵敏度和准确性。结果表明，在标准试验条件下，在590 nm处的吸光度测量和450 nm的比例是严格蛋白质浓度的线性。这个简单的程序，提高了精度和提高约10倍，允许定量下降到50吴牛血清白蛋白的检测灵敏度。此外，常用的洗涤剂，用于创建的细胞裂解液引入的干扰大大减少了新的协议。线性方程的集体行动和比尔定律的基础上开发的，完全符合实验数据。

Watch this Scientific Journal Video about Linearization of the Bradford Protein Assay at JoVE.com

Linearization of the Bradford Protein Assay

受到损害的内在非线性的快速，便捷的Bradford法测定蛋白质的准确性和灵敏度。我们展示一个简单的的线性过程，大大提高了准确性，提高约10倍的检测灵敏度，并显着减少洗涤剂干扰。

Bradford蛋白含量的线性

Determination of microgram quantities of protein in the Bradford Coomassie brilliant blue assay is accomplished by measurement of absorbance at 590 nm. ...

Research

JoVE Journal

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The actin cytoskeleton within the cell is a network of actin filaments that allows the movement of cells and cellular processes, and that generates ...

科研

生物学

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Ubiquitination, the covalent attachment of the polypeptide ubiquitin to target proteins, is a key posttranslational modification carried out by a set of ...

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In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents ...

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Investigation of gene function in diverse organisms relies on knowledge of how the gene products interact with each other in their normal cellular ...

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In this article, a simple, quantitative, liquid phase affinity capture assay is presented. Provided that one protein can be tagged and another protein ...

Linking Predation Risk, Herbivore Physiological Stress and Microbial Decomposition of Plant Litter

链接捕食风险，草食动物生理应激和植物凋落物的微生物分解

The quantity and quality of detritus entering the soil  determines the rate of decomposition by microbial communities as well as  recycle rates of ...

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Protein degradation by the ubiquitin-proteasome system (UPS) is a major regulatory mechanism for protein homeostasis in all eukaryotes. The standard ...

Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes

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Centrosomes are small but important organelles that serve as the poles of mitotic spindle to maintain genomic integrity or assemble primary cilia to ...

Identifying Protein-protein Interaction Sites Using Peptide Arrays

鉴定蛋白质相互作用的网站使用多肽阵列

Protein-protein interactions mediate most of the processes in the living cell and control homeostasis of the organism. Impaired protein interactions may ...