
CD34 functions as a cell-cell adhesion factor and cell-surface glycoprotein. It may also mediate the attachment of stem cells to bone marrow extracellular matrixes or directly to stromal cells. Cells expressing CD34 are normally found in the umbilical cord and bone marrow as hematopoletic cells, and in vascular endothelium. In addition to stem cell recognition, CD34 is expressed by vascular endothelium; it appears that proliferating endothelial cells express this molecule in greater amounts than resting cells. In comparison to factor VIII R Antigen, CD34 stains are stronger and appear to be more sensitive in nature.
In tumors, CD34 is found in Alveolar Soft Part Sarcoma, pre B-ALL (positive in 75%), AML(40%), AMLM7 (most), Dermatofibrosarcoma Protuberans, Gastrointestinal Stromal Tumors, Giant Cell Fibroblastoma, Granulocytic Sarcoma, Kaposi’s Sarcoma, Liposarcoma, Malignant Fibrous Histiocytoma, Malignant Peripheral Nerve Sheath tumors, Mengingeal Hemangiopericytomas, Meningiomas, Neurofibromas, Schwannomas, and Papillary Thyroid Carcinoma. A negative CD34 may exclude Ewing’s Sarcoma/PNET, Myofibrosarcoma of the breast, and Inflammatory Myofibroblastic tumors of the stomach.
For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).
CD34 is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.
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- Aziza J, et al. Am J Clin Pathol. 1990;96:25-31
Recommended Immunohistochemical Protocol
- Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
- Air dry for 2 hours at 58° C.
- Deparaffinize, dehydrate and rehydrate tissues.
- Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
- Any of three heating methods may be used:
- Electric Pressure Cooker
Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature. - Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes. - Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
- Electric Pressure Cooker
- After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
- Wash slides with IHC wash buffer or DI water.
- Continue IHC staining protocol.
Immunohistochemical Protocol
ImmunoDetector
(AP or HRP)
(AP or HRP)