mAbs Against HUMAN Protein
BCL6
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | ST42B |
| PROTEIN: | human full length Bcl-6 |
| PROTEIN WEB: | http://www.hprd.org/summary?hprd_id=00180&isoform_id=00180_3&isoform_name=Isoform_1 |
| ANTIGEN USED: | GST-BCL6 recombinant protein |
| 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: | IHQ-frozen, IHQ-paraffin, WB, IF |
| APP NO TESTED: | Flow Cytometry, IP |
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
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 | ST42B | 1:4 | supernatant | None | goat anti mouse DAKO | Tonsil / | nuclear | mouse | |
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | ST42B | 1:4 | supernatant | 20 min R2 (Tris-EDTA) | Novolink kit | Tonsil / | nuclear | mouse | Available |
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | ST42B | neat | supernatant | Tris-EDTA | goat anti mouse ALEXA | Tonsil / | nuclear | mouse | Available |
Enlarge image
- BCL6 antibody (ST42B) in HEK transfected cells.
- Validation of ST42B monoclonal antibody in transfected cells. Nuclear and cytoplasmic staining on frozen cytospin preparations of transfected HEK293T-BCL6 cells using antibody ST42B. Hek-v5-CSF1R transfected cells were used as negative control.
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- 42B mAb staining in human frozen tonsil.
- In tonsil, BCL6 protein is strongly expressed by germinal center B cells
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- ST42B 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.
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- Triple immunofluorescence staining.
- BCL6 (ST42B) nuclear protein in blue, PD-1 membrane staining in red and CD 57 membrane staining in green.
| 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 | ST42B | neat | supernatant | Tonsil / | 79kDa | mouse | Available | |
| Immunoprecipitation | ||||||||
Not tested Not tested | / | mouse | ||||||
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- Western Blotting of BCL6 (ST42B) using different cell extracts.
- WB analysis of cell line samples and human tonsil extract for Bcl-6.
Anti-tubulin was used as loading control.
Lane 1 Ramos cell line (100ug) (+)
Lane 2 Colo205 cell line (100ug) (-)
Lane 3 Akata cell line (100ug) (+)
Lane 4 Karpas 299 cell line (100ug) (+)
Lane 5 Toledo cell line (100ug) (+)
Lane 6 Tonsil (100ug) (+)
Lane 7 Hut78 (100ug) (-)