CD34 (CD34) Antikörper

Antigen: CD34 (CD34)

Klonalität: Monoklonal (QBEnd-10)

Wirt: Maus

Reaktivität: Human

Applikation: Immunhistochemie (Paraffinschnitte) (IHC (p))

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370720

Produktbeschreibung

Weitere Bezeichnung: CD34 Class II

Immunogen: Endothelial cell membranes obtained as vesicles from human placenta (6, 8).

Isotyp: IgG1, kappa chain  (Passende Sekundärantikörper)

Klon: QBEnd-10

Beschreibung: CD34 is a single-chain transmembrane protein of approximately 116 kDa. CD34 is expressed on immature haematopoietic stem/progenitor cells, capillary endothelial cells, embryonic fibroblasts and rare glial cells in nervous tissue (1, 4). CD34 appears to be expressed at its highest level on the earliest progenitors, and to decrease progressively with maturation (2, 3). CD34 is a stage-specific, rather than a lineage-specific, leucocyte differentiation antigen. The most immature definable B-lymphoid precursors (CD19+/CD10+/TdT+) are CD34+ (2-4). Immature T-lymphoid precursors also express TdT and CD34 (4). Normal peripheral blood lymphocytes, monocytes, granulocytes, and platelets do not express CD34. Approximately 60% of acute B-lymphoid leukaemias and 40% of acute myeloid leukaemias (AML), and 1% to 5% of acute T-lymphoid leukaemias express CD34 (3). Chronic lymphoid leukaemias, lymphomas and multiple myelomas have been found to be uniformly CD34 negative (1-4). Monoclonal antibodies to CD34 can be confined to three main classes, class I, class II and class III, defined by the differential sensitivity of the corresponding CD34 epitopes to degradation by specific enzymes. The QBEnd 10 antibody is a class II monoclonal antibody that recognizes a CD34 epitope that is resistant to neuraminidase, and sensitive to glycoprotease and chymopapain (1, 3, 7).

Spezifität: Anti-CD34, QBEnd 10, was included in the Fourth and Fifth International Workshops and Conferences on Human Leucocyte Differentiation Antigens (Vienna 1989, Boston 1993), and studies by different laboratories confirmed its reactivity with the CD34 antigen (2, 7) and the class II epitope (7). In formalin-fixed, paraffin embedded tonsil sections the antibody labels endothelial cells. In flow cytometry the antibody labels KG-1a cells (a primitive myeloid leukaemia cell line, known to be CD34+).

Anwendungen

Applikationshinweise: Paraffin sections: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pre-treatment of tissues with proteinase K, trypsin, or heat-induced epitope retrieval is required. For heat-induced epitope retrieval, optimal results are obtained with, Dako Target Retrieval Solution, High pH, code S3308, or 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0. Less optimal results are obtained with Dako Target Retrieval Solution, code S1700, or 10 mmol/L citrate buffer, pH 6.0. The tissue sections should not dry out during the treatment or during the following immunocytochemical staining procedure. Frozen sections and cell preparations: The antibody can be used for labelling acetone fixed, frozen sections and cell smears. Dilution: Monoclonal Mouse Anti-Human CD34 Class II, code No. M7165, may be used at a dilution range of 1:25-1:50 when applied on formalin-fixed, paraffin-embedded sections of human tonsil and using 20 minutes heat-induced epitope retrieval in 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0, and 30 minutes incubation at room temperature with the primary antibody. Optimal conditions may vary depending on specimen and preparation method, and should be determined by each individual laboratory. The recommended negative control is Dako Mouse IgG1, code X0931, diluted to the same mouse IgG concentration as the primary antibody. Unless the stability of the diluted antibody and negative control has been established in the actual staining procedure, it is recommended to dilute these reagents immediately before use, or dilute in Dako Antibody Diluent, code S0809. Positive and negative controls should be run simultaneously with patient specimen. Visualization: Dako LSAB/HRP kit, code K0679, and Dako EnVision/HRP kits, code K4004 and K4006, are recommended. For frozen sections and cell preparations, the Dako APAAP kit, code K0670, is a good alternative if endogenous peroxidase staining is a concern. Follow the procedure enclosed with the selected visualization kit. Automation: The antibody is well suited for immunocytochemical staining using automated platforms, such as the Dako Autostainer. Cells labelled by the antibody display staining confined to the cell surface membrane. Normal tissues: The antibody labels capillaries of most tissues as well as umbilical artery and, to a lesser extent, vein. The antibody does not label endothelium of most large vessels and the endothelium of placental sinuses (8). Abnormal tissue: A total of 112 vascular tumours, 54 carcinomas of different types and 45 non-vascular spindle cell tumours were stained with anti-CD34, clone QBEnd-10 (6). All (22/22) the benign tumours of blood vascular origin showed immunoreactivity whereas only five out of eight lymphangiomas demonstrated a weak focal reaction with QBEnd-10 (6). Tumour cells in angiosarcoma forming vasoformative areas and solid areas showed immunopositivity to QBEnd-10 in 17/23 and 13/24 cases respectively (6). Proliferating vessels and the majority of spindle cells in Kaposi's sarcoma were positive in all 40 cases. Only one of 54 cases of carcinoma showed luminal reaction to QBend-10. However, 17 of 45 spindle cell tumours displayed a positive reaction (6). Product-specific limitations It has been reported that binding of CD34 monoclonal antibodies may be reduced by fixatives present in erythrocyte-lysing reagents, suggesting that the CD34 epitope is negatively affected. Hence, fixatives should be used with caution if the monoclonal antibody is to be used for enumeration purposes (9).

Buffer: Monoclonal mouse antibody provided in liquid form as cell culture supernatant dialysed against 0.05 mol/L Tris/HCl, pH 7.2, and containing 15 mmol/L NaN 3 ,

Lagerung: Store at 2-8 °C. Precautions: 1.For professional users. 2. This product contains sodium azide (NaN 3 ), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used. 4. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin. 5. Unused solution should be disposed of according to local, State and Federal regulations

Forschungsgebiet: CD Antigene, Oberflächenrezeptoren der Immunzellen

Beschränkungen: Für Forschungszwecke. Als CE-zertifizierter Antikörper in der Europäischen Union für In Vitro Diagnostik (IVD) zugelassen.

Publikationen

Publikationen: Gerecht, Ferreira, Langer: "Vascular differentiation of human embryonic stem cells in bioactive hydrogel-based scaffolds." in: Methods in molecular biology (Clifton, N.J.), Vol. 584, pp. 333-54, 2009 (PubMed).

McCarthy, OBrien, Cummins et al.: "GIST with a twist--upregulation of PDGF-B resulting in metachronous gastrointestinal stromal tumor and dermatofibrosarcoma protuberans." in: Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract, Vol. 14, Issue 2, pp. 398-403, 2010 (PubMed).

Arvidsson, Bergstroem, Arvidsson et al.: "Hypoxia stimulates CXCR4 signalling in ileal carcinoids." in: Endocrine-related cancer, Vol. 17, Issue 2, pp. 303-16, 2010 (PubMed).

Hansen, Suorensen, Spindler et al.: "Microvessel density and the association with single nucleotide polymorphisms of the vascular endothelial growth factor receptor 2 in patients with colorectal cancer." in: Virchows Archiv : an international journal of pathology, Vol. 456, Issue 3, pp. 251-60, 2010 (PubMed).