Evaluation of glycoprotein Ib expression on feline platelets
| Autor: | Fern Tablin, Naomi J. Walker, Jocelyn D. Johnsrude |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Blood Platelets
Immunoprecipitation Enzyme-Linked Immunosorbent Assay Flow cytometry chemistry.chemical_compound medicine Animals Platelet Fluorescent Antibody Technique Indirect Gel electrophoresis General Veterinary biology medicine.diagnostic_test Leupeptin Antibodies Monoclonal Calpain General Medicine Actin cytoskeleton Flow Cytometry Molecular biology Precipitin Tests Specific Pathogen-Free Organisms Biochemistry Glycoprotein Ib chemistry Platelet Glycoprotein GPIb-IX Complex biology.protein Cats |
| Zdroj: | American journal of veterinary research. 62(2) |
| ISSN: | 0002-9645 |
| Popis: | Objective—To determine whether platelets obtained from cats expressed glycoprotein Ib (GPIb). Sample Population—Platelets obtained from 11 specific-pathogen-free cats. Procedure—Platelets were analyzed by use of immunofluorescence microscopy, flow cytometry, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, western immunoblot analysis, and immunoprecipitation. Results—Immunofluorescence microscopy and flow cytometry revealed the protein on the surface of feline platelets. Biochemical studies (western immunoblot analysis and immunoprecipitation) revealed a 140-kd membrane glycoprotein. Additional biochemical studies revealed that feline GPIb was sensitive to proteolysis, because platelet cytoskeletons prepared with low concentrations of a calpain inhibitor (ie, leupeptin; 100 µg/ml) had substantial proteolysis, and there was an association of protein fragments with the actin cytoskeleton. Conclusions and Clinical Relevance—Analysis of these results indicate that feline platelets express a 140-kd membrane protein that is recognized by monoclonal antibodies developed against GPIb. Application of standardized ELISA to quantitate glycocalicin, the water-soluble fragment of GPIb, may provide important information on the production of microvesicles, increased platelet turnover, and abnormal proteolysis. (Am J Vet Res 2001;62:195–201) |
| Databáze: | OpenAIRE |
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