Cytokeration 7 Antibody Staining Protocol for Immunohistochemistry


Description: Cytokeratins are intermediate filament cytoskeletal proteins essential to development and differentiation of epithelial cells. Approximately twenty different cytokeratins have been identified and are classified and numbered according to molecular weight and isoelectric points. In general, most low molecular weight cytokeratins (40-54 kDa) are distributed in nonsquamous epithelium, Moll’s catalog numbers 7-8 and/or 17-20. High molecular weight cytokeratins (48-67 kDa) are found in the squamous epithelium, Moll’s catalog numbers 1-6 and/or 9-16. Cytokeratin 7 is one of the basic type II cytokeratins and is found in most glandular epithelia and in transitional epithelia. The cytokeratin has been identified as cytokeratin 7 according to the classification of Moll et al. Monoclonal mouse anti-human Cytokeratin 7, Clone OV-TL 12/30 (anti-CK7) is to identify epithelial cells in normal and neoplastic tissues using immunohistochemical methods. Anti-CK7 is valuable as a histodiagnostic tool for the subtyping of various carcinomas and tumors of epithelial origin. Anti-CK7 may provide a more precise classification of many epithelial tumors whose differential diagnosis is otherwise difficult.


Primary Antibody

Name: Cytokeratin 7 Antibody

Clone: OV-TL 12/30, Mouse anti-Human

Supplier: Dako

Catalog Number: M7018

Dilution: 1:300 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

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: 95-100 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic/membrane
Images: Search image

Additional Information:
Tissue Type: Lung, ovary, cervix, endometrium, and breast, skin.
Fixation: Formalin fixed paraffin sections, acetone fixed frozen sections, or cell smears.
Positive Control: Lung, ovary, cervix, endometrium, and breast, 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


There is no need to perform antigen retrieval on frozen sections or cell smears