A simple method for quantitative, semiquantitative, and qualitative assay of protein

✍ Scribed by Asim Esen

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 588 KB
• Volume: 89
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90749-2

✦ Synopsis

A simple protein determination method is described which gives a linear response from 0.05 to 3-4 mg/ml (or more) of protein and is suitable for dilute (<0.05 mglml) protein solutions if concentrated on paper by repeated spotting. Five-microliter solutions of a standard protein and the unknown are spotted on Whatman No. 1 chromatography paper and stained with 0.1% Coomassie brilliant blue R-250. After destaining with water and drying, the stained spots are cut with a cork borer. The dye-protein complex is eluted with 1% SDS, and the absorbance of dye-protein complex is measured at 600 nm. This assay is specific for proteins and virtually free from interference by common laboratory reagents and other nonproteinaceous substances. The color of dye-protein complex is stable for months. There is no specific period of time required for the assay. It can be stopped at a given step, then completed at a more convenient time.