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mAbs Against HUMAN Protein

FOXP3

Antibody Validated
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
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
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
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.