mAbs Against HUMAN Protein
BCL11AXL
| STATUS: |  |
|---|---|
| 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 | 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 | 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.