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mAbs Against HUMAN Protein

BCL7A

Antibody Validated
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
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
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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).
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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
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) (+)
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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) (-)