One-step HPLC purification procedure for porcine brain 90-kDa heat shock protein
| Autor: | Wen-Chuan Lee, Lawrence Fu-nien Chu, Teh-yang Huang, Simon J.T. Mao, Redman Chu, Ping-Cheng Yang |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Gel electrophoresis
Brain Chemistry Chromatography Chemistry medicine.drug_class Swine Sodium Immunoblotting chemistry.chemical_element Chemical Fractionation Monoclonal antibody High-performance liquid chromatography Silver stain Blot Biochemistry Heat shock protein Protein purification medicine Animals Electrophoresis Gel Two-Dimensional Electrophoresis Polyacrylamide Gel HSP90 Heat-Shock Proteins Chromatography High Pressure Liquid Biotechnology Densitometry |
| Zdroj: | Protein expression and purification. 10(2) |
| ISSN: | 1046-5928 |
| Popis: | The 90-kDa heat shock protein (HSP90) was purified from porcine brain by a novel single-step purification procedure using diethylaminoethyl high-performance liquid chromatography (HPLC). About 4.8 mg of HSP90 was isolated from 25 g wet wt porcine brain tissue. The purified protein possessed a single moiety on one- and two-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis followed by silver staining. Western blotting using monoclonal antibody prepared against human HSP90 confirmed its identity as HSP90. These results indicate that small-scale HPLC purification of HSP90 from porcine brain tissue can be readily accomplished, with high yield, using a convenient one-step purification method. The procedure described in this paper represents a significant improvement in current purification methods for the isolation of HSP90 from porcine brain. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect