Detection of myosin heavy chain in skeletal muscles using monoclonal antibodies on formalin fixed, paraffin embedded tissue sections
| Autor: | Nancyleigh E. Carson, Jiang Gu, C. David Ianuzzo |
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| Předmět: |
Histology
biology medicine.drug_class medicine.medical_treatment Skeletal muscle Proteinase K Monoclonal antibody Molecular biology Staining Medical Laboratory Technology medicine.anatomical_structure Myosin biology.protein medicine Immunohistochemistry Anatomy Cardiomyoplasty Immunostaining Biomedical engineering |
| Zdroj: | Web of Science |
| Popis: | To meet the demand for an easy and accurate method to identify fast and slow skeletal muscle fiber types in the ongoing clinical trial of cardiomyoplasty, we developed protocols of tissue preparation and immunohistochemical staining using monoclonal antibodies to MHC-I and II. A range of enzymatic and antigen recovery solutions were tested, and an optimal procedure was established for each antibody. Enzyme digestion with 20 μg/ml of proteinase K at 42°C for 30 min was most suited to unveiling MHC-I and Target Unmasking Fluid gave the best results for MHC-II. The staining results were comparable to conventional ATPase staining and immunostaining on unfixed cryostat sections. The modified procedures used in a double irnmunostaining protocol allowed simultaneous demonstration and distinction of the 2 fiber types on the same tissue section. This approach provided the centers, participating in the ongoing cardiomyoplasty clinical trial, a uniform and easy method to assess muscle fiber types and facili... |
| Databáze: | OpenAIRE |
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