Influence of detergents on determination of heat shock protein 70 by ELISA.
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概要
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To objectively evaluate the relationship between psychiatric disorders and stress, we attempted to measure the levels of 70 kilodalton (kDa) heat shock proteins (HSP70) in peripheral blood mononuclear cells (PBMC) using ELISA. Although the standard curve created from measurements of a commercial HSP70 (from bovine brain; S-HSP70) showed good linearity, measurements of HSP70 in PBMC samples appeared negative when concentrations were calculated using this curve. We determined that addition of the detergent used for PBMC lysis to dilutions of S-HSP70 also resulted in negative calculations. Therefore, when we dissolved the PBMC in distilled water instead of the detergent, HSP70 levels could be correctly determined. To verify the accuracy of HSP70 calculations for the PBMC samples dissolved in distilled water, we compared the ELISA results with those obtained by SDS-polyacrylamide gel electrophoresis and Western blotting for PBMC dissolved in the detergent. Both methods resulted in similar patterns of HSP70 expression.
- 日本電気泳動学会の論文
日本電気泳動学会 | 論文
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