 Methods & Techniques For Histologists and Immunohistochemists |
c-kit Oncoprotein (CD117) Antibody Staining Protocol for Immunohistochemistry
Description: The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor, c-kit (CD117) which is closely related to the platelet derived growth factor family. c-kit plays a role during haematopoiesis, gametogenesis and melanogenesis.
Primary Antibody
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Name: c-kit Oncoprotein (CD117) Antibody |
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Clone: 57A5D8, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-cKIT |
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Dilution: 1:200 - 1:400 |
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Incubation Time/Temp: 60 minutes/room temperature |
Antigen Retrieval
| Device: No antigen retrieval is needed |
| Buffer/pH value: N/A |
| Heat/Cool Temperature: N/A |
| Heat/Cool Time: N/A |
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: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane/cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Skin |
| Fixation: Acetone fixed frozen sections. |
| Positive Control: Normal skin, epidermal melanocytes. |
| 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 paraffin sections |
References: