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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
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Name: CD117 (c-Kit) Antibody |
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Clone: 2B8, Rat anti-Mouse |
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Supplier: Research Diagnostic |
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Catalog Number: RDI-MCD117-2B8 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
Notes:
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There is no need to perform antigen retrieval for acetone fixed frozen sections |
References: