CD117 (c-Kit) Antibody Staining Protocol for Immunohistochemistry


Description: This antibody reacts with CD117 which in adult bone marrow is expressed on hematopoietic progenitor cells including CD90, Ter-119-CD45R/B220- CD11b, Ly-6G, CD4-CD8-Sca-1 multipotent hematopoietic stem cells, progenitors committed to myeloid and/or erythroid precursors of B and T lymphocytes. Clone 2B8 does not block the action of c-kit


Primary Antibody

Name: CD117 (c-Kit) Antibody

Clone: 2B8, Rat anti-Mouse

Supplier: Research Diagnostic

Catalog Number: RDI-MCD117-2B8

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: 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: Cell surface staining
Images: Search image

Additional Information:
Tissue Type: Bone marrow
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Bone marrow
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