Bcl-2 Oncoprotein antibody

Antigen: Bcl-2 Oncoprotein

Clonality: Monoclonal (124)

Host: Mouse

Reactivity: Human

Application: Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Certificates: ISO 9001:2008, ISO 13485:2003

Supplier: Dako

Catalog no.: ABIN370622

Additional Information

Alternative name: BCL2 Oncoprotein

Immunogen: Synthetic peptide comprising amino acids 41-54 of human BCL2 oncoprotein (1).

Isotype: IgG1, kappa  (Matching secondary antibodies)

Clone: 124

Description: BCL2 oncoprotein is a blocker of apoptotic cell death. Gene transfer experiments have shown that elevated levels of this protein can protect a wide variety of cells from diverse cell death stimuli ranging from growth factor withdrawal and cytotoxic lymphokines to virus infection and DNA-damaging, anticancer drugs and radiation (2, 3). BCL2 oncoprotein resides on the cytoplasmic side of the mitochondrial outer membrane, endoplasmic reticulum and nuclear envelope (2, 4), and has a molecular mass of 26 kDa (3). The BCL2 gene is involved in the t(14,18) chromosomal translocation found in 85% of human follicular lymphomas and 20% of diffuse B-cell lymphomas (4). In this translocation, the BCL2 gene at chromosome segment 18q21 is juxtaposed with the Ig heavy chain locus at 14q32, resulting in deregulated expression of BCL2 oncoprotein (4).

Specificity: In Western blotting of extracts of normal human spleen (1, 5), t(14,18)-positive follicular lymphoma (1), and myloid leukaemic cell lines (5) the antibody labels solely a band of 26 kDa, corresponding to BCL2 oncoprotein under both nonreducing (1) and reducing conditions (1, 5). The antibody labels the myeloid leukaemic cell lines, HL-60 (promyelocytic), KG1 (myeloblastic), GM-1 (monoblastic) and K562 (erythromyeloid) (5).

Application Details

Application Notes: Paraffin sections: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin or Bouin's (5). Pre-treatment of tissues with heat-induced epitope retrieval is required. For tissues fixed in formalin, optimal results are obtained with 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0. Less optimal results are obtained with Dako Target Retrieval Solution, High pH, code S3308, Dako Target Retrieval Solution, code S1700, or 10 mmol/L citrate buffer, pH 6.0. Pre-treatment of tissues with proteinase K was found inefficient. The tissue sections should not dry out during the treatment or during the following immunohistochemical staining procedure. Frozen sections and cell preparations: The antibody can be used for labelling acetone-fixed, frozen sections and cell smears (1). Dilution: Monoclonal Mouse Anti-Human BCL2 Oncoprotein, code M0887, may be used at a dilution range of 1:50-1:100 when applied on formalin-fixed, paraffin-embedded sections of tonsil tissue 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, codes 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. Performance characteristics: Cells labelled by the antibody display a cytoplasmic staining pattern. Normal tissues: The antibody labels almost all peripheral blood lymphocytes. In lymphoid tissue, small lymphocytes in the mantle zones and T-cell areas are positive whereas very few cells in germinal centres are labelled. In the spleen, many cells in both T- and B-cell areas and the red pulp are labelled by the antibody. In the thymus, many cells in the medulla are labelled, while most cells in the cortex show weak or negative staining (1).

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 .

Storage: 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.

Restrictions: For in vitro diagnostic use

Images

Follicular lymphoma. The majority of the neoplastic cells showa distinct cytoplasmic staining reaction.

Burkitt lymphoma/leukemia. The neoplastic cells are negativefor BCL2 and only benign T cells are stained.

Tonsil. The scattered T cells in the germinal centers and themantle zone B cells show a moderate to strong cytoplasmic stainingreaction. The germinal center B cells are negative.

Publications

Publications: Frid, Shekhonin, Koteliansky et al.: "Phenotypic changes of human smooth muscle cells during development: late expression of heavy caldesmon and calponin." in: Developmental biology, Vol. 153, Issue 2, pp. 185-93, 1992 (PubMed).

Glukhova, Frid, Koteliansky: "Developmental changes in expression of contractile and cytoskeletal proteins in human aortic smooth muscle." in: The Journal of biological chemistry, Vol. 265, Issue 22, pp. 13042-6, 1990 (PubMed).

Sartore, De Marzo, Borrione et al.: "Myosin heavy-chain isoforms in human smooth muscle." in: European journal of biochemistry / FEBS, Vol. 179, Issue 1, pp. 79-85, 1989 (PubMed).

Borrione, Zanellato, Scannapieco et al.: "Myosin heavy-chain isoforms in adult and developing rabbit vascular smooth muscle." in: European journal of biochemistry / FEBS, Vol. 183, Issue 2, pp. 413-7, 1989 (PubMed).

Lazard, Sastre, Frid et al.: "Expression of smooth muscle-specific proteins in myoepithelium and stromal myofibroblasts of normal and malignant human breast tissue." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, Issue 3, pp. 999-1003, 1993 (PubMed).

Savera, Gown, Zarbo: "Immunolocalization of three novel smooth muscle-specific proteins in salivary gland pleomorphic adenoma: assessment of the morphogenetic role of myoepithelium." in: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, Vol. 10, Issue 11, pp. 1093-100, 1997 (PubMed).

Vergis, Corbishley, Thomas et al.: "Expression of Bcl-2, p53, and MDM2 in Localized Prostate Cancer with Respect to the Outcome of Radical Radiotherapy Dose Escalation." in: International journal of radiation oncology, biology, physics, 2010 (PubMed).

Tabrizi, Kalloger, Koebel et al.: "Primary ovarian mucinous carcinoma of intestinal type: significance of pattern of invasion and immunohistochemical expression profile in a series of 31 cases." in: International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists, Vol. 29, Issue 2, pp. 99-107, 2010 (PubMed).

Papaxoinis, Triantafyllou, Sasco et al.: "Subsite-specific differences of estrogen receptor beta expression in the normal colonic epithelium: implications for carcinogenesis and colorectal cancer epidemiology." in: European journal of gastroenterology & hepatology, Vol. 22, Issue 5, pp. 614-9, 2010 (PubMed).

Hoogwater, Nijkamp, Smakman et al.: "Oncogenic K-Ras Turns Death Receptors Into Metastasis-Promoting Receptors in Human and Mouse Colorectal Cancer Cells." in: Gastroenterology, 2010 (PubMed).

Salerni, Bates, Albershardt et al.: "Vinblastine induces acute, cell cycle phase-independent apoptosis in some leukemias and lymphomas and can induce acute apoptosis in others when mcl-1 is suppressed." in: Molecular cancer therapeutics, Vol. 9, Issue 4, pp. 791-802, 2010 (PubMed).