Cytokeration - Basal Cell Antibody Staining Protocol for Immunohistochemistry

 

Description: Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in epithelial tissues by at least 20 different polypeptides. They range in molecular weight from between 40 kDa and 68 kDa and isoelectric pH between 4.9 - 7.8, The individual cytokeratin polypeptides are designated 1-20. The various epithelia in the human body usually express cytokeratins, which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. MAB3224 is a broadly reacting cytokeratin antibody which reacts with cytokeratin 5 among others. It stains basal cells in combined and stratified epithelial tissues. It recognizes the stem cell population, including the so-called amplifying cell in the prostate epithelium.

 

Primary Antibody

Name: Cytokeratin, Basal Cell Antibody

Clone: Mouse monoclonal

Supplier: Chemicon

Catalog Number: MAB3224

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: Stomach
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Stomach
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: