The characterization and use of different antibodies against the hsp70 major heat shock protein family for the development of an immunoassay
| Autor: | Peter V. Nacharov, Michael Welsh, Boris A. Margulis, Alexander V. Kinev, Olga I. Tsvetkova |
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| Rok vydání: | 1991 |
| Předmět: |
Antiserum
Immunoassay medicine.diagnostic_test medicine.drug_class Clinical Biochemistry Immunoblotting Enzyme-Linked Immunosorbent Assay Biology Monoclonal antibody Biochemistry Molecular biology Precipitin Tests Epitope Analytical Chemistry Antigen Polyclonal antibodies Antibody Specificity medicine biology.protein Humans Denaturation (biochemistry) Antibody Heat-Shock Proteins HeLa Cells |
| Zdroj: | Electrophoresis. 12(9) |
| ISSN: | 0173-0835 |
| Popis: | The hsp70 family of major stress proteins is composed of several different members exhibiting similar structural and functional properties. In order to obtain an antiserum with wide epitope reactivity, rabbits were immunized with a mixture of native and denatured hsp70 purified from bovine muscle by ATP-affinity chromatography. Screening for antibody specificity was performed by a "sandwich" enzyme linked immunosorbent assay (ELISA). Immunoprecipitation and immunoblotting analyses demonstrated that the polyclonal antiserum obtained by us and a monoclonal antibody raised against a different preparation of antigen recognized the same determinant on the native hsp70 molecule (inducible form). With a different specificity the polyclonal antiserum recognized only the denatured monomers of the other members of the hsp70 family. These results are discussed in relation to the immunological features of the hsp70 molecule and to the development of an immunoassay for the detection of hsp70 in cell and tissue extracts. |
| Databáze: | OpenAIRE |
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