mAbs Against HUMAN Protein
BCL6
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | GI191E |
| PROTEIN: | human full length BCL6 |
| PROTEIN WEB: | http://www.hprd.org/summary?hprd_id=00180&isoform_id=00180_3&isoform_name=Isoform_1 |
| ANTIGEN USED: | pcDNA3-BCL6 |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | human and mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
| APP RECOMMENDED: | WB, IHC-frozen, IHC-paraffin |
| APP NO TESTED: | IP, IF, Flow Cytometry |
Description
BCL-6 is a transcriptional regulator gene which codes for a 706-amino-acid nuclear zinc finger protein. Antibodies to this protein stain the germinal centre cells in lymphoid follicles, the follicular cells and interfollicular cells in Follicular Lymphoma, Diffuse Large B-Cell Lymphomas, and Burkitt’s lymphoma, and the majority of the Reed-Sternberg cells in Nodular Lymphocyte Predominant Hodgkin’s Disease. In contrast, anti-BCL-6 rarely stains Mantle Cell Lymphoma and MALT Lymphoma. BCL-6 expression is seen in approximately 45% of CD30+ Anaplastic Large Cell Lymphoma but is consistently absent in other peripheral T-cell lymphomas.
Publication describing antibody characterization/validation
García JF, García JF, Maestre L, Lucas E, Sánchez-Verde L, Romero-Chala S, Piris MA, Roncador G. Genetic Immunization: a New Monoclonal Antibody for the Detection of BCL-6 Protein in Paraffin Sections. J Histochem Cytochem. 2006 54(1):31-8. http://www.ncbi.nlm.nih.gov/pubmed/16046671
References
Guggisberg K and Jordan R. Mantle cell lymphoma of the oral cavity: case series and comprehensive review of the literature. Oral and maxillofacial pathology. 2010 109 (1): 98-104.
Kanellis G, Roncador G, Arribas A, Mollejo M, Montes-Moreno S, Maestre L, Campos-Martin Y, Rıos Gonzalez JR, Martinez-Torrecuadrada JL, Sanchez-Verde L, Pajares R, Cigudosa JC, Martin MC and Piris MA. Identification of MNDA as a new marker for nodal marginal zone lymphoma. Leukemia 2009 23, 1847–1857.
Maestre L, Tooze R, Cañamero M, Montes-Moreno S, Ramos R, Doody G, Boll M, Barrans S, Baena S, Piris MA and Roncador G. Expression pattern of XBP1(S) in human B-cell lymphomas. Haematologica. 2009 March; 94(3): 419–422.
Montes-Moreno S, Roncador G, Maestre L, Martínez N, Sánchez-Verde L, Camacho FI, Cannata J, Martínez-Torrecuadrada JL, Shen Y, Chan W and Piris MA. Gcet1 (centerin), a highly restricted marker for a subset of germinal center-derived lymphomas. Blood. 2008 111: 351-358.
Garcia JF, Roncador G, Garcia JF, Sanz AI, Maestre L, Lucas E, Montes-Moreno S, Fernandez Victoria R, Martinez-Torrecuadrada JL, Marafioti T, Mason DY, Piris MA. PRDM1/BLIMP-1 expression in multiple B and T-cell lymphoma. Haematologica. 2006. Apr; 91(4):46.
García JF, García JF, Maestre L, Lucas E, Sánchez-Verde L, Romero-Chala S, Piris MA, Roncador G. Genetic Immunization: a New Monoclonal Antibody for the Detection of BCL-6 Protein in Paraffin Sections. J Histochem Cytochem. 2006 54(1):31-8.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | GI191E | 1:100 | supernatant | None | goat anti mouse DAKO | Tonsil / | nuclear | ||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | GI191E | 1:350 | supernatant | 20 min R2 (Tris-EDTA) | Novolink kit | Tonsil / | nuclear | mouse | Available |
| Immunofluorescence | |||||||||
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- GI191EmAb in transfected cells
- Nuclear and cytoplasmic staining on frozen cytospin preparations of transfected HEK293T/BCL6 cells using antibody GI191E. HEK-a-myb transfected cells were used as negative control.
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- GI191E mAb in frozen tonsil section
- In tonsil, BCL6 protein is strongly expressed by germinal center B cells
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- GI191E staining in human paraffin sections.
- In tonsil, BCL6 protein is strongly expressed by germinal center B cells. Testicle section was used as a negative control.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | GI191E | supernatant | 1:20 | Raji cell line / | 78kDa | 80kDa | Available | |
| Immunoprecipitation | ||||||||
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- Western Blotting of BCL6 (GI191E) using different human cell lines.
- Lane 1 Raji cell line (50ug) (+)
Lane 2 U937 cell line (50ug) (+)
Lane 3 HL-60 cell line (50ug) (+)
Lane 4 k562 cell line (42ug) (+)
Lane 5 HeLa cell line (50ug) (+)