Cytokeratin Antibody Staining Protocol for Immunohistochemistry


Description: The cytokeratins (CKs) belong to the intermediate filaments, which create a cytoskeleton in almost all eukaryotic cells. In contrast to other intermediate filaments, CKs are made up of a highly complex multigene family of polypeptides with molecular masses ranging from 40 to 68 kDa. CKs are generally held to belong to the most fundamental markers of epithelial differentiation, and until now, 20 distinct CK polypeptides have been revealed in various human epithelia. The CKs can be divided into an acidic type A (class I) and a neutral-basic type B (class II) subfamily.


Primary Antibody

Name: Cytokeratin Antibody

Clone: MNF116, Mouse anti-Human

Supplier: Dako

Catalog Number: M0821

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.

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: 95-100 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
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Skin
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


Not need to perform antigen retrieval on acetone fixed frozen sections