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Actin (Muscle) Antibody Staining Protocol for Immunohistochemistry
Description: This antibody reacts with the muscle actin on formalin fixed paraffin embedded sections or frozen sections.
Primary Antibody
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Name: Mouse Anti-Human Actin, Muscle |
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Clone: HHF35 |
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Supplier: Dako |
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Catalog Number: M0635 |
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Dilution: 1:200 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: Antigen retrieval is not needed |
| Buffer/pH value: N/A |
| Heat/Cool Temperature: N/A |
| Heat/Cool Time: N/A |
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: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane/Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin-fixed paraffin sections or acetone fixed frozen sections |
| Positive Control: Skeletal muscle, tongue, leg muscle |
| 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 |
Notes:
There is no need to perform antigen retrieval on acetone fixed frozen sections |
References: