 Methods & Techniques For Histologists and Immunohistochemists |
Actin (alpha Smooth Muscle) Antibody Staining Protocol for Immunohistochemistry
Description: This antibody reacts with the alpha-smooth muscle isoform of actin present in smooth muscle cells of vessels and different parenchymes, myoepithelial cells, pericytes and some stromal cells in the intestine, testis and ovary. It does not react with actin from fibroblasts (beta-and gamma-cytoplasmic), striated muscle (alpha-sarcomic), and myocardium (alpha-myocardial).
Primary Antibody
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Name: Mouse Anti-Human Actin, alpha Smooth Muscle |
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Clone: 1A4 |
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Supplier: Dako |
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Catalog Number: M0851 |
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Dilution: 1:1000 |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: Steamer |
| Buffer/pH value: Citrate buffer/pH 6.0 |
| Heat/Cool Temperature: 95 - 100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane/Cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Smooth muscle, colon |
| Fixation: Formalin-fixed paraffin sections or acetone fixed frozen sections as well as cell smears. |
| Positive Control: Smooth muscle, colon |
| Negative Control: Omit primary antibody, isotype control, absorption control |
| Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
References: