CD117 (c-Kit) Antibody Staining Protocol for Immunohistochemistry


Description: The stem cell factor receptor, c-kit (CD117), acts as an inhibitor of apoptotic cell death upon binding of its ligand. Strong positive immunoreactivity is observed in mast cells of epithelial and hematopoietic tissues. Glial cells from both fetal and adult brain show positive staining, also ductal cells in mammary glands. Tumor cells:c-Kit expression is demonstrated in small cell and large cell lung carcinoma.


Primary Antibody

Name: CD117 (c-Kit) Antibody

Clone: Rabbit polyclonal

Supplier: Dako

Catalog Number: A4502

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: 60min/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:
Tissue Type: Human tonsil, intestinal tissues, skin, brain, lung
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Human tonsil, intestinal tissues, skin, brain, 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


There is no need to perform antigen retrieval for acetone fixed frozen sections