Ultramicro-analysis by Use of Light-Scanning Photoacoustic Densitometry for Electrophoresed Protein in Human Hair
✍ Scribed by Toshiro Fukami; Katsumi Uchiyama; Yoshihiro Yoshimura; Takaho Watanabe; Hiroyuki Nakazawa
- Publisher
- Elsevier Science
- Year
- 1996
- Tongue
- English
- Weight
- 120 KB
- Volume
- 238
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
sured indirectly using a photoacoustic (PA) 2 method A technique was developed for the ultramicro-analy- (1). Quantitative methods using a light-scanning PA sis of proteins electrophoresed by Laemmli's method densitometer combined with dye or silver staining have using a light-scanning photoacoustic densitometer. been previously reported (2, 3). When electrophoresed After electrophoresis, the proteins were blotted on a gels are measured by the PA method, gels are usually nitrocellulose membrane filter and colored by the aviused after drying to suppress the effect of water. Howdin-biotin complex method. This filter was then meaever, protein bands on gels are easily distorted due to sured using a photoacoustic densitometer. The optithe changes in gels upon drying. In addition, gels are mal blotting time was 150 min. The relative standard difficult to store. On the other hand, Masujima et al. deviation of four measurements was 3.89% for 200 ng used a PA method for immunocomplex detection after bovine serum albumin (BSA). The limits of detection trapping on a membrane filter followed by coloration were 2.3, 0.69, 4.4, and 2.9 ng (S/N Å 3) for BSA, oval-(4). Therefore, we studied ultramicro-detection using a bumin, carbonic anhydrase, and a-Lactoalbum respeclight-scanning PA densitometer for proteins separated tively. Proteins eluted from hair by various harmful on gels by SDS-PAGE (5). This method was applied agents, such as a surfactant and UV irradiation, were to the quantitative analysis of proteins eluted from huanalyzed by the present method. Using 2-mercaptoethman hair. Hair was damaged by various treatments, anol, the molecular weights of proteins in hair were in such as UV irradiation and permanent wave lotion