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Cytokeratin 7 Antibody Staining Protocol for Immunohistochemistry
Description: The antibody reacts with cytokeratin 7 which has been found in columnar and glandular epithelium of the lung, cervix, breast, bile ducts and larger collecting ducts of the kidney. Occasionally staining of blood vessel cell walls, particularly endothelial cells, may be observed.
Primary Antibody
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Name: Cytokeratin 7 FITC conjugate Antibody |
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Clone: LP5K, Mouse anti-Human |
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Supplier: Chemicon |
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Catalog Number: CBL194F |
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Dilution: 1:50 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) or IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
| 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, lung, breast, kidney. |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Skin, lung, breast, kidney. |
| 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: