Cytokeration 8&18 Antibody Staining Protocol for Immunohistochemistry

 

Description: Cytokeratins are a family of water-insoluble intermediate filaments, composed of polypeptide chains found in the cytoskeleton of epithelial cells. Twenty different forms of human cytokeratins exist (Moll numbers 1-20) in both normal and malignant epithelial cell lines.  Specific combinations of cytokeratin peptides are associated with certain epithelial cells, and are therefore useful in the characterization of poorly differentiated carcinoma. Cytokeratins may also be clinically significant indicators in distinguishing carcinoma from malignant tumors of non-epithelial origin, such as lymphoma, sarcoma, and melanoma. Zymed’s Zym5.2 antibody detects human cytokeratins with molecular weights 52 and 45 kDa, corresponding to cytokeratins 8 and 18, respectively. This antibody is useful in the detection of adenocarcinomas, esotheliomas, and certain squamous epithelia-derived carcinomas. Interestingly, some epithelial tumors demonstrate cytokeratin 8 and 18 expression, but normal stratified squamous epithelium does not. Cytokeratin 8 and 18 staining has also been observed in renal cell carcinomas, hepatocellular carcinomas, cholangiocarcinomas, germ cell tumors, and some neuroendocrine carcinomas, epithelioid sarcomas, and non-epithelial tumors.

 

Primary Antibody

Name: Cytokeratin 8&18 Antibody

Clone: UCD/PR-10.11, Mouse monoclonal

Supplier: Zymed Labs

Catalog Number: 08-1213

Dilution: Pre-dilute

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval

Device: Steamer

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/membrane

Images: Search image

Additional Information:

Tissue Type: Skin, pancreas

Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections.

Positive Control: Skin, pancreas

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: