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mAbs Against HUMAN Protein

BCL11AXL

Antibody Validated
CONTACT INFORMATION:LRF Haemato-oncology Group, University of Oxford
STATUS:Validated
TYPE:mouse anti human
CLONE NAME:BCL11A/123
PROTEIN:C-terminus of extra long isoform of BCL11A, BCL11AXL (aa 637-835)
PROTEIN WEB:http://www.ncbi.nlm.nih.gov/omim/606557
ANTIGEN USED:Bacterially expressed GST-fusion protein of human BCL11AXL
FUSION PARTNER:NS1
ISOTYPE:IgG1
SPECIES REACTIVITY:Human
PREPARATION AND STORAGE:Aliquot and store at 4oC. Do not freeze.
APP RECOMMENDED:IHQ-paraffin, IHQ-frozen, WB
APP NO TESTED:IF, IP, Flow cytometry

DESCRIPTION

BCL11A is a Krüppel zinc-finger transcription factor identified originally from a chromosome translocation in aggressive B-cell chronic lymphocytic leukaemia. The gene may function as both an oncogene and a tumour suppressor. Alternative splicing generates multiple isoforms, the BCL11AXL being the largest and most abundant transcript. The protein is differentially expressed during B-cell development and is strongly expressed in plasmacytoid dendritic cells. The BCL11A/123 antibody specifically recognises the extra long form of the protein and not the long or short forms of BCL11A. The BCL11A/123 antibody is not cross reactive with the long form of the BCL11B protein that shares significant sequence identity with the immunogen.

REFERENCES

Pulford K, Banham AH, Lyne L, et al. The BCL11AXL transcription factor: its distribution in normal and malignant tissues and use as a marker for plasmacytoid dendritic cells. Leukemia. 2006;20:1439-1441.

Weniger MA, Pulford K, Gesk S, et al. Gains of the proto-oncogene BCL11A and nuclear accumulation of BCL11A(XL) protein are frequent in primary mediastinal B-cell lymphoma. Leukemia. 2006;20:1880-1882.

Liu H, Ippolito GC, Wall JK, et al. Functional studies of BCL11A: characterization of the conserved BCL11A-XL splice variant and its interaction with BCL6 in nuclear paraspeckles of germinal center B cells. Mol Cancer. 2006;5:18.

Marafioti T, Paterson JC, Ballabio E, et al. Novel markers of normal and neoplastic human plasmacytoid dendritic cells. Blood. 2008;111:3778-3792.

Chetaille B, Bertucci F, Finetti P, et al. Molecular profiling of classical Hodgkin lymphoma tissues uncovers variations in the tumor microenvironment and correlations with EBV infection and outcome. Blood. 2009;113:2765-3775

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
BCL11A/123 neat supernatant Dako Envision Tonsil / COS1 Nucleus NT
Paraffin tissue
Recommended Result obtained is satisfactory. The reagent can be use in this application
BCL11A/123 1/10 supernatant Tris/EDTA Dako Envision Tonsil / COS1 Nucleus NT
Immunofluorescence
Enlarge image
Immunolabelling of BCL11AXL transfected COS1 cells
Nuclear staining of frozen COS/ BCL11AXL transfectants but not BCL11AL transfectants using antibody BCL11A/123.
Enlarge image
Tonsil staining of BCL11AXL with BCL11A/123
BCL11AXL is present in tonsillar B cells (brown) in germinal centres and mantle zones. Double labelling studies show the strongly labelled cells in the interfollicular areas to be CD123-positive plasmacytoid dendritic cells (arrowhead).
WB Techniques
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
BCL11A/123 neat supernatant tonsil / COS1 91kDa approx 120kDa NT
Immunoprecipitation
Enlarge image
Western blotting of BCL11 transfectants
Western Blotting characterization of BCL11A/123 monoclonal antibody with lysates of transfectants expressing different BCL11A isoforms and the highly homologous BCL11B protein. Shows specificity of the BCL11A/123 antibody for the detection of BCL11AXL.