BCL6 Protein Antikörper

Antigen: BCL6 Protein

Klonalität: Monoklonal (PG-B6p)

Wirt: Maus

Reaktivität: Human

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

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370732

Produktbeschreibung

Immunogen: Recombinant glutathione S-transferase (GST)-BCL6 protein corresponding to amino acids 3-484 of the human BCL6 protein (1).

Isotyp: IgG1, kappa chain  (Passende Sekundärantikörper)

Klon: PG-B6p

Beschreibung: The proto-oncogene BCL6 encodes a 92-98 kDa POZ/zinc finger protein. Through its six C-terminal zinc-finger motifs, the BCL6 protein binds to specific DNA sequences of target genes, where it acts as a transcriptional repressor (2, 3). BCL6 expression may be ubiquitous, as low levels are detected in many tissues. However, high levels of BCL6 expression are limited to cells of the B-lymphoid linage and skeletal muscle. In the B-lymphoid cell linage, BCL6 expression seems to be tightly regulated during differentiation, and strong expression is observed preferentially in germinal centre (GC) B cells, but not in plasma cells, and precursor, mantle zone, and memory B cells. Neoplastic counterparts of GC B cells, including Burkitt’s, follicular, and diffuse large cell lymphomas, also display high levels of BCL6 expression (3). BCL6 may be implicated in chromosomal translocations where the regulatory region of the BCL6 gene is replaced by a heterologous reciprocal partner such as the immunoglobulin (IG) genes. Promotor substitution leads to deregulation of the BCL6 expression, which may be associated with lymphomagenesis (3). BCL6 rearrangement is one of the most common genetic abnormalities in B-cell non-Hodgkin’s lymphoma, with an especially high frequency (28-40%) in diffuse large cell lymphoma (2).

Spezifität: SDS-PAGE analysis of immunoprecipitates formed between the antibody and total lysates of Bjab cells (BCL6+) and BCL6 transfected EB3 cells, shows reaction with a ~95 kDa fragment corresponding to BCL6. No reaction is observed with Rd cells (BCL6-) and EB3 cells transfected with a control plasmid (1). In Western blotting of total lysates of Bjab cells and BCL6 transfected EB3 cells, or control transfected EB3 cells, the antibody did not label any bands, suggesting lack of reactivity with denatured BCL6 protein (1). As demonstrated by immunocytochemistry, the antibody cross-reacts with the BCL6-equivalent protein in various mammals, including cow, rabbit, rat, sheep, and swine. No cross-reactivity was observed in chicken and pigeon (1).

Anwendungen

Applikationshinweise: Paraffin sections: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pre-treatment of tissues with heat-induced epitope retrieval is required. 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. However, Dako Target Retrieval Solution, code S1700, or 10 mmol/L citrate buffer, pH 6.0 and pre-treatment of tissues with proteinase K were 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. Dilution: Monoclonal Mouse Anti-Human BCL6 Protein, code M7211, may be used at a dilution range of 1:10-1:20 when applied on formalin-fixed, paraffin-embedded sections of human tonsil and using 20 minutes heat-induced epitope retrieval in Dako Target Retrieval Solution, High pH, code S3308, 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 immunohistochemical staining using automated platforms. Cells labelled by the antibody show a diffuse/microgranular staining confined to the nucleus.

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.

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

Bilder

Diffuse large B-cell lymphoma (germinal center subtype). Themajority of the neoplastic cells show a distinct nuclear staining reaction.

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

Tonsil. The majority of the germinal center B cells show amoderate to strong nuclear staining reaction.

Publikationen

Publikationen: Alacacioglu, Ozcan, Ozkal et al.: "Prognostic significance of immunohistochemical classification of diffuse large B-cell lymphoma." in: Hematology (Amsterdam, Netherlands), Vol. 14, Issue 2, pp. 84-9, 2009 (PubMed).

Obermann, Csato, Dirnhofer et al.: "BCL2 gene aberration as an IPI-independent marker for poor outcome in non-germinal-centre diffuse large B cell lymphoma." in: Journal of clinical pathology, Vol. 62, Issue 10, pp. 903-7, 2009 (PubMed).

Pinto, Andrue, Silva et al.: "BCL-6 oncoprotein in breast cancer: loss of expression in disease progression." in: Pathobiology : journal of immunopathology, molecular and cellular biology, Vol. 76, Issue 5, pp. 235-42, 2009 (PubMed).

Dojcinov, Venkataraman, Raffeld et al.: "EBV positive mucocutaneous ulcer--a study of 26 cases associated with various sources of immunosuppression." in: The American journal of surgical pathology, Vol. 34, Issue 3, pp. 405-17, 2010 (PubMed).