Cytokeration 18 Antibody Staining Protocol for Immunohistochemistry

 

Description: The antibody reacts with a large number of epithelial cell types including many ductal and glandular epithelia. In general, it does not react with squamous epithelium. Mixed epithelia of glands consist of two distinct cell types where cytokeratin 8 and 18 are expressed in the luminal cells. Like all cytokeratins, cytokeratin 18 is, in general, not expressed by cells of non-epithelial origin. The antibody does not recognize other intermediate filament proteins.

 

Primary Antibody

Name: Cytokeratin 18 Antibody

Clone: DC10, Mouse anti-Human

Supplier: Dako

Catalog Number: M7010

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.

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

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Skin
Fixation: Formalin fixed paraffin sections, acetone fixed frozen sections, or cytospin preparations.
Positive Control: Skin, kidney, lung
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:

There is no need to perform antigen retrieval for frozen sections.

 

References: