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-Tek^TM 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-Tek^TM Epitope Retrieval Steamer Set (Cat# IW-1102)

Buffer/pH value: IHC-Tek^TM 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: 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

Notes:

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

References: