Microassay of proteins on membrane filter in the nanogram range using cycloheptaamylose-dansyl chloride complex
✍ Scribed by Toshio Kinoshita; Fumio Iinuma; Akio Tsuji
- Publisher
- Elsevier Science
- Year
- 1975
- Tongue
- English
- Weight
- 365 KB
- Volume
- 66
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A protein assay is described in which the sample is spotted on a Sartorius membrane filter and dansylated with cycloheptaamylose-dansyl chloride complex dissolved in aqueous urea solution. The fluorescence intensity of the spot on the membrane was directly measured by a scanning fluorometer. This assay is rapid, simple, highly reproducible, and linear from 0.05 to 4 pg of protein. Very dilute solutions of protein can also be determined by filtering off the sample on the membrane filter in the presence of 0.1 M MgCl, and dansylating the protein collected on the membrane. There is no interference either by commonly used reagents such as Tris, citrate, guanidine, and glycine, or by naturally occurring amines and amino acids.
1 This paper constitutes part V of the series entitled "Microanalysis of Proteins and Peptides"; part IV: T. Kinoshita, F. Iinuma, A. Tsuji and I. Moriguchi (1974), Chem. Pharm.