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CD34 Antibody Staining Protocol for Immunohistochemistry
Description: Recognizes a single chain transmembrane glycoprotein of 110kDa, which is identified as CD34. This antigen is selectively expressed on human lymphoid and myeloid hematopoietic progenitor cells. The antibody to CD34 also reacts with vascular endothelial cells in normal tissues, and in benign and malignant proliferations. The utility of CD34 has value in the studies of benign and malignant vacular tumors as will as characterization of acute leukemia in bone marrow. CD34 has been used to measure angiogenesis in many types of tumors which reportedly predicts tumor recurrence. It is also used to differentiate dermatofibrosarcoma protuberans from fibrous histiocytoma.
Primary Antibody
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Name: CD34 Antibody |
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Clone: QBEnd/10, Mouse anti-Human |
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Supplier: Biocare Medical |
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Catalog Number: CM084B |
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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: 60 min/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: Membrane/Cell surface |
| Images: Search image |
Additional Information:
| Tissue Type: Skin, tonsil |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Skin, tonsil, angiosarcoma |
| Negative Control: Omit primary antibody, isotype control, 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 need to perform antigen retrieval on acetone fixed frozen sections |
References: