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Cytokeratin 19 Antibody Staining Protocol for Immunohistochemistry
Description: Cytokeratin polypeptide 19 (CK19) is a type I intermediate filament protein that is expressed in stratified and simple-type epithelia. Cytokeratins 8 and 19 concentrate at costameres of striated muscle and copurify with the dystrophin-glycoprotein complex, perhaps through the interaction of the cytokeratins with the actin-binding domain of dystrophin. Studies in COS-7 cells and in vitro showed that dystrophin binds directly and specifically to K19. Results show that dystrophin interacts through its actin-binding domain with CK19 specifically and are consistent with the idea that cytokeratins associate with dystrophin at the sarcolemma of striated muscle. CK19 is synthesized mainly in embryonic and adult simple epithelia, but has also been found in stratified epithelia as well. CK19 is the smallest known keratin and is remarkable in that, contrary to all other keratins, it does not have a designated partner for the formation of filaments, implying that regulation of its expression is different from other keratin-encoding genes.
Primary Antibody
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Name: Cytokeratin 19 (Krt 19) |
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Clone: Rabbit monoclonal |
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Supplier: Epitomics |
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Catalog Number: 3863-1 |
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Dilution: 1:100 |
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Incubation Time/Temp: 60 min/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/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: Membrane/cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Mouse |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Mouse colon tissue |
| 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: