mAbs Against HUMAN Protein
FOXP3
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | 221D |
| PROTEIN: | Human full length FOXP3 |
| ANTIGEN USED: | GST-FOXP3 recombinant protein |
| FUSION PARTNER: | myeloma p3-NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | human, mouse, cat, dog, horse, rabbit, alpaca and pig. |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
| APP RECOMMENDED: | IHQ-paraffin, IHQ-frozen, WB, Flow cytometry, IF |
| APP NO TESTED: | IP |
Description
FOXP3 is a forkhead-family transcription factor that negatively regulates T cell function. Mice carrying a loss-of-function mutation in FoxP3 present with fatal autoimmune-like disease caused by hyperresponsive CD4(+) T cells. Mice that overexpress scurfin possess fewer mature T cells with reduced functional capabilities compared with normal littermate control mice. FOXP3 is critical for normal CD4+ T cell function and for the successful coordination of a normal response to immunological challenge in vivo.
Publication describing antibody characterization/validation
Roncador G, Brown PJ, Maestre L, Hue S, Martínez-Torrecuadrada JL, Ling KL, Pratap S, Toms C, Fox BC, Cerundolo V, Powrie F, Banham AH. Analysis of FOXP3 protein expression in human CD4(+)CD25(+) regulatory T cells at the single-cell level. Eur J Immunol 2005 35:1681-1691. http://www.ncbi.nlm.nih.gov/pubmed/15902688
Banham AH, Lyne L, Scase TJ, Blacklaws BA. Monoclonal antibodies raised to the human FOXP3 protein can be used effectively for detecting Foxp3(+) T cells in other mammalian species. Vet Immunol Immunopathol. 2009 Feb 15;127(3-4):376-81. http://www.sciencedirect.com/science/article/pii/S0165242708007083
References
Roncador G, Brown PJ, Maestre L, Hue S, Martínez-Torrecuadrada JL, Ling KL, Pratap S, Toms C, Fox BC, Cerundolo V, Powrie F, Banham AH. Analysis of FOXP3 protein expression in human CD4(+)CD25(+) regulatory T cells at the single-cell level. Eur J Immunol 2005 35:1681-1691.
Banham AH, Lyne L, Scase TJ, Blacklaws BA. Monoclonal antibodies raised to the human FOXP3 protein can be used effectively for detecting Foxp3(+) T cells in other mammalian species. Vet Immunol Immunopathol. 2009 Feb 15;127(3-4):376-81.
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 | 221D | 1:2 | supernatant | Tonsil / | nuclear | mouse | |||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 221D | 1:20 | supernatant | 20 min ER2 (Tris-EDTA) | Novolink kit | Tonsil / | nuclear | mouse | Available |
| Immunofluorescence | |||||||||
Enlarge image
- Anti-FOXP3 antibody in transfected cells
- Hek-V5-MNDA was used as negative control.
Enlarge image
- FOXP3 (221D) expression on human frozen tonsil.
Enlarge image
- Anti-FOXP3 antibody on human paraffin sections.
| 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 | 221D | supernatant | neat | MT2 cell line / Kidney | 47kDa | 47kDa | Available | |
| Immunoprecipitation | ||||||||
Enlarge image
- Western Blotting characterization of 221D monoclonal antibody
- Western Blotting characterisation of 221D monoclonal antibody
Antibody 221D recognises HEK-FOXP3 transfected cells and MT-2 cell line (human T cell leukemia virus carrier cell line) by WB.
Lane1 HEK-FOXP3 transfected cells (20ug) (+)
Lane 2 HEK-mTIMP2 transfected cells (20ug) (-)
Lane 3 MT2 cell line (150ug) (+)
Lane 4 Human tonsil (150ug) (-)
Lane 5 Human kidney (150ug) (-)
| OTHERS | Title | Description | Protocol |
|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | Epitope map of anti-FOXP3 (221D) antibody |
Enlarge image
- Epitope map of anti-FOXP3 (221D) antibody
- The figure shows Foxp3 protein functional domains and the location of epitope recognized by 221D antibody (aa105-236). Foxp3 contains a Zn finger/Leucine zipper and a conserved Forkhead Domain.