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

Name: CD34 Antibody

Clone: QBEnd/10, Mouse anti-Human

Supplier: Biocare Medical

Catalog Number: CM084B

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: 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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


Not need to perform antigen retrieval on acetone fixed frozen sections