首页   按字顺浏览 期刊浏览 卷期浏览 Protein concentration by precipitation with pyrogallol red prior to electrophoresis
Protein concentration by precipitation with pyrogallol red prior to electrophoresis

 

作者: Thomas Marshall,   Nicola J. Abbott,   Peter Fox,   Katherine M. Williams,  

 

期刊: ELECTROPHORESIS  (WILEY Available online 1995)
卷期: Volume 16, issue 1  

页码: 28-31

 

ISSN:0173-0835

 

年代: 1995

 

DOI:10.1002/elps.1150160107

 

出版商: Wiley Subscription Services, Inc., A Wiley Company

 

关键词: Protein concentration;Pyrogallol red;Urinary proteins

 

数据来源: WILEY

 

摘要:

AbstractThe pyrogallol red protein assay (Clinical Chemistry1986,32, 1551–1554) is based upon formation of a blue protein‐dye complex in the presence of molybdate under acidic conditions. However, centrifugation of the assay mixture results in loss of color yield and precipitation of the protein‐dye complex which can be recovered and resolubilized to achieve protein concentration prior to sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The method has been evaluated relative to trichloroacetic acid (TCA) precipitation for recovery and electrophoresis of commercial protein and peptide molecular weight markers. Precipitation with pyrogallol red‐molybdate (PRM) gives better and more uniform recovery of both proteins and peptides as compared to TCA. The lower limit of PRM precipitation is similar to TCA and corresponds to 1 μg protein per mL assay mixture. This is equivalent to 100 μL of 10 μg/mL protein using the standard protein assay or 1 μg/mL protein using a modified assay incorporating a fivefold concentrate of the dye reagent. Application of the method is demonstrated by concentration of urinary proteins. The method is simple and economic and useful for conserving trace amounts of precious sample as it allows recovery of protein for electrophoresis following

 

点击下载:  PDF (434KB)



返 回