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CD34

IHC of CD34 on an FFPE Tonsil Tissue

Description

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.

Antibody Type
Mouse Monoclonal
Clone
QBEnd/10
Isotype
IgG1
Reactivity
Paraffin, Frozen
Localization
Membranous
Control
Tonsil, Placenta, Appendix
Storage
Store at 2°-8°C
Stability
2 years

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).

Presentation

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.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 5225
Prediluted
Ready-To-Use
3.0 ml
BSB 5226
Prediluted
Ready-To-Use
7.0 ml
BSB 5227
Prediluted
Ready-To-Use
15.0 ml
BSB 5228
Concentrated
1:25-1:100
0.1 ml
BSB 5229
Concentrated
1:25-1:100
0.5 ml
BSB 5230
Concentrated
1:25-1:100
1.0 ml
BSB 5231
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Civin CL, et al. London Academic Press. 1989:818-825
  2. Fina L, et al. Blood. 1990;75:2417-2426
  3. Ramani P, et al. Histopathology. 1990;17:237-242
  4. Aziza J, et al. Am J Clin Pathol. 1990;96:25-31

Recommended Immunohistochemical Protocol

Pretreatment
  1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
  2. Air dry for 2 hours at 58° C.
  3. Deparaffinize, dehydrate and rehydrate tissues.
  4. 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).
  5. 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.
  6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
  7. Wash slides with IHC wash buffer or DI water.
  8. Continue IHC staining protocol.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain