Ultramicro-analysis by use of light-scanning photoacoustic densitometry for electrophoresed protein in human hair.
A technique was developed for the ultramicro-analysis of proteins electrophoresed by Laemmli's method using a light-scanning photoacoustic densitometer. After electrophoresis, the proteins were blotted on a nitrocellulose membrane filter and colored by the avidin-biotin complex method. This filter was then measured using a photoacoustic densitometer. The optimal blotting time was 150 min. The relative standard deviation of four measurements was 3.89% for 200 ng bovine serum albumin (BSA). The limits of detection were 2.3, 0.69, 4.4, and 2.9 ng (S/N = 3) for BSA, ovalbumin, carbonic anhydrase, and alpha-Lactoalbum respectively. Proteins eluted from hair by various harmful agents, such as a surfactant and UV irradiation, were analyzed by the present method. Using 2-mercaptoethanol, the molecular weights of proteins in hair were in the range of 14,000-64,000. Maximal elution of protein was obtained at pH 8. More protein was eluted under alkaline conditions than under acidic conditions. A protein of Mr 68,000 was eluted from hair by oxidative treatment with either UV irradiation or sodium bromate.
Fukami
T
T
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Hoshi University, 2-4-41, Tokyo, Shinagawa-ku, 142, Japan.
Uchiyama
K
K
Yoshimura
Y
Y
Watanabe
T
T
Nakazawa
H
H
D000162:Acoustics; D001959:Bromates; D002138:Calibration; D003720:Densitometry / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006197:Hair / Q000737:chemistry* / Q000528:radiation effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008027:Light; D008832:Microchemistry / Q000379:methods*; D010084:Oxidation-Reduction; D011506:Proteins / Q000032:analysis*; D017670:Sodium Compounds; D013501:Surface-Active Agents; D014466:Ultraviolet Rays
D001959:Bromates; D011506:Proteins; D017670:Sodium Compounds; D013501:Surface-Active Agents; C060553:sodium bromate