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Cytokeratin 10 Antibody Staining Protocol for Immunohistochemistry
Description: Cytokeratins comprise a diverse group of intermediate filament proteins (IFPs) that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. Cytokeratins have been found to be useful markers of tissue differentiation which is directly applicable to the characterization of malignant tumors. Cytokeratins 10 and 13 are present in the cytoskeletal region of a subset of squamous cell carcinomas. Cytokeratin 10 is a heterotetramer of two type I and two type II keratins, is generally associated with keratin 1, and is seen in all suprabasal cell layers including stratum corneum.
Primary Antibody
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Name: Cytokeratin 10 (RKSE60) |
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Clone: Monoclonal, Mouse anti-Human |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-23877 |
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Dilution: 1:100 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 of epithelial tissue |
| Images: Search image |
Additional Information:
| Species reactivity: Human, rat, mouse, canine. |
| Fixation: Formalin fixed paraffin sections. |
| Positive Control: Skin. |
| 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:
Not tested on frozen sections. |
References: