Cytokeratin 10 Antibody Staining Protocol for Immunohistochemistry

 

Description: Cytokeratins comprise a diverse group of intermediate filament proteins (IFPs) that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. Cytokeratins have been found to be useful markers of tissue differentiation which is directly applicable to the characterization of malignant tumors. Cytokeratins 10 and 13 are present in the cytoskeletal region of a subset of squamous cell carcinomas. Cytokeratin 10 is a heterotetramer of two type I and two type II keratins, is generally associated with keratin 1, and is seen in all suprabasal cell layers including stratum corneum.

 

Primary Antibody

Name: Cytokeratin 10 (RKSE60)

Clone: Monoclonal, Mouse anti-Human

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-23877

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 ºC - 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 of epithelial tissue
Images: Search image

Additional Information:
Species reactivity: Human, rat, mouse, canine.
Fixation: Formalin fixed paraffin sections.
Positive Control: 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

Notes:

Not tested on frozen sections.

 

References: