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Desmin Antibody Staining Protocol for Immunohistochemistry
Description: The antibody reacts with desmin from human, pig, rat and taod. In tissue sections this antibody is used to stain skeletal, cardiac, visceral, and some vascular smooth muscle cells. Cell lines such as RD (ATCC CCL 136) and hamster BHK-21 are positive.
Primary Antibody
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Name: Desmin Antibody |
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Clone: DE-B-5, Mouse monoclonal |
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Supplier: Chemicon |
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Catalog Number: MAB3430 |
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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 minutes/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95 ºC - 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: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Skin, skeletal muscle, cardiac tissue (heart), GI tract (stomach, intestine, colon). |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Skin, skeletal muscle, cardiac tissue (heart), GI tract (stomach, intestine, colon). |
| Negative Control: Omit primary antibody, isotype control or 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:
No need to perform antigen retrieval on frozen sections |
References: