mAbs Against HUMAN Protein
BCL7A
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | 15C |
| PROTEIN: | Bcl7a human full length |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/omim/601406 |
| ANTIGEN USED: | His-GST-Bcl7a recombinant protein |
| FUSION PARTNER: | myeloma p3-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-paraffin, IF |
| APP NO RECOMMENDED: | WB, IHQ-frozen |
| APP NO TESTED: | IP, Flow cytometry |
Description
This gene is directly involved, with Myc and IgH, in a three-way gene translocation in a Burkitt lymphoma cell line. As a result of the gene translocation, the N-terminal region of the gene product is disrupted, which is thought to be related to the pathogenesis of a subset of high-grade B cell non-Hodgkin lymphoma.
Publication describing antibody characterization/validation
BCL7A protein expression in normal and malignant lymphoid tissues. Ramos-Medina R, Montes-Moreno S, Maestre L, Cañamero M, Rodríguez-Pinilla M, Martínez-Torrecuadrada J, Piris MÁ, Majid A, Dyer MJ, Pulford K, Roncador G. Br J Haematol. 2013 Jan;160(1):106-9.
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 | 15C | 1:20 | supernatant | goar anti mouse HRP DAKO | Tansil / | nuclear | mouse | ||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 15C | 1:40 | supernatant | Tris-EDTA | ER2 20min Novolink | Tonsil / | nuclear | mouse | Available |
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 15C | 1:20 | supernatant | tris-EDTA | Tonsil / | nuclear | mouse | Available | |
Enlarge image
- BCL7A antibody (15C) in transfected cells
- Nuclear and cytoplasmic staining of BCL7A mAb in transfected HEK293-BCL7A cells. No reactivity with BCL7 family members Bcl7b, Bcl7c was found.
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- 15C staining in human lymphoid tissue
- BCL7A is expressed in the nuclei of germinal center (GC) B cells and in mantle cells. Some cells in the interfollicular area are also BCL7A positives. Spleen showing BCL7a staining of GC B cells, mantle cells and marginal zone lymphocytes (weak staining) (b). Normal thymus shows Bcl7a staining mainly in T-cells in the cortex, however, some BCL7A positive cells can also be found in the medulla (c).
Enlarge image
- BCL7A (15C) immunofluorescence in human paraffin tonsil
- BCL7A (15C) nuclear protein in red, ki67 nuclear staining in green and DAPI staining in blue.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Inconclusive Result obtained is not completely satisfactory. More testing are needed before using these reagents in this application | 15C | neat | supernatant | raji / l540 | 26kDa | 30kDa | Available | |
| Immunoprecipitation | ||||||||
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- WB of Bcl7a (clone15C) mAB
- Western Blotting using cell extracts of cells transfected with BCL7A, BCL7B, and BCL7C expression vectors. A 35 kDa band was detected in HEK293T BCL7A cell extract but was absent in HEK293T BCL7B and BCL7C transfectants.
Lane 1 Hek-Bcl7a trasfected cells (20ug) (+)
Lane 2 Hek-Bcl7b transfected cells (20ug) (-)
Lane 3 Hek-Bcl7c transfected cells (20ug) (-)
Lane 4 Bcl7a-HIS recombinant protein (0.1ug) (+)
Enlarge image
- WB of 15C mAn in human cell lines
- Antibody 15C is not able to detect BCL7A endogenous protein by WB.
Predicted molecular weight: 26kDa
Observed molecular weight: 31kDa
Lanes
Lane 1 Transfected Hek-pcdna3-V5-Bcl7a (20цg) (+)
Lane 2 Transfected Hek-pcdna3-Bcl7b (20цg) (-)
Lane 3 Transfected Hek-pcdna3-Bcl7c (20цg) (-)
Lane 4 Hela (200цg) (-)
Lane 5 DOHH2 (200цg) (+)
Lane 6 L540 (200цg) (-)
Lane 7 Raji (200цg) (-)