ARG54202
anti-FOXP3 antibody [3G3]
anti-FOXP3 antibody [3G3] for Flow cytometry,Western blot and Human,Mouse
Cell Biology and Cellular Response antibody; Gene Regulation antibody; Immune System antibody; Regulatory T cells Study antibody
Overview
| Product Description | Mouse Monoclonal antibody [3G3] recognizes FOXP3 |
|---|---|
| Tested Reactivity | Hu, Ms |
| Tested Application | FACS, WB |
| Specificity | The mouse monoclonal antibody 3G3 recognizes Nterminal region of FoxP3, a 4755 kDa transcription factor, which is the master regulator in the development and function of regulatory T cells. |
| Host | Mouse |
| Clonality | Monoclonal |
| Clone | 3G3 |
| Isotype | IgG1 |
| Target Name | FOXP3 |
| Antigen Species | Mouse |
| Immunogen | Full-length His-tagged recombinant murine FoxP3 |
| Conjugation | Un-conjugated |
| Alternate Names | XPID; AIID; JM2; IPEX; PIDX; Forkhead box protein P3; Scurfin; DIETER |
Application Instructions
| Application Suggestion |
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| Application Note | FACS: Staining method: - Perform staining of cell surface markers (CD25, CD4 etc.) for 10 min. at 4°C in the dark; 1 million cells in 100 µl final volume. - Add 1-2 ml of PBS with 1% BSA, centrifugate at 300g and discard the supernatant. Further steps perform on ice and with ice-cold reagents. - Resuspend the cells in 1 ml of cold fixation solution (Miltenyi Biotec) and incubate for 30 min. on ice. - Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. - Resuspend the cells in 1-2 ml of ice-cold PBS with 1% BSA. - Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. - Resuspend the cells in 1-2 ml of ice-cold permeabilization solution (Miltenyi Biotec) and incubate 5 min. - Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. - Resuspend the cells in 80µl of ice-cold permeabilization solution and add 20 µl of FcR blocking solution and incubate for 5 min. (4°C, in the dark). - Perform intracellular staining of FoxP3 for 30 min. (4°C, in the dark) with appropriate amount of 3G3 antibody (final concentration 2-10 µg/ml; optimal concentration for peripheral blood is 3 µg/ml). - Add 1-2 ml of ice-cold permeabilization solution and incubate for 5 min. at 4°C in the dark. - Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. - Resuspend the cells in 300 µl of ice-cold PBS with 1% BSA and measure on a FACS device with appropriate setting as soon as possible. Keep cold until measuring. (If necessary, for the same sample use mouse IgG1 isotype control (MOPC-21) in the same format as anti-FoxP3 antibody 3G3.) * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. |
Properties
| Form | Liquid |
|---|---|
| Purification | Purified from ascites by protein-A affinity chromatography. |
| Purity | > 95% (by SDS-PAGE) |
| Buffer | PBS (pH 7.4) and 15 mM Sodium azide |
| Preservative | 15 mM Sodium azide |
| Concentration | 1 mg/ml |
| Storage Instruction | For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C or below. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. |
| Note | For laboratory research only, not for drug, diagnostic or other use. |
Bioinformation
| Database Links | |
|---|---|
| Gene Symbol | Foxp3 |
| Gene Full Name | forkhead box P3 |
| Background | FoxP3 (Forkhead box protein 3), a highly conserved forkhead/winged-helix transcription factor, plays a crucial role in maintaining immune homeostasis by governing the development and function of regulatory T cells. It is constitutively expressed at high level in CD25+ CD4+ Treg cells and at low level in a CD25- CD4+ Treg cell subset. Defects in gene encoding FoxP3 protein cause the scurfy phenotype in mice, and in human the IPEX syndrome (immune dysfunction, polyendocrinopathy, enteropathy, X-linked syndrome), also known as X-linked autoimmunity-allergic dysregulation (XLAAD) syndrome. |
| Function | Transcriptional regulator which is crucial for the development and inhibitory function of regulatory T-cells (Treg). Plays an essential role in maintaining homeostasis of the immune system by allowing the acquisition of full suppressive function and stability of the Treg lineage, and by directly modulating the expansion and function of conventional T-cells. Can act either as a transcriptional repressor or a transcriptional activator depending on its interactions with other transcription factors, histone acetylases and deacetylases. The suppressive activity of Treg involves the coordinate activation of many genes, including CTLA4 and TNFRSF18 by FOXP3 along with repression of genes encoding cytokines such as interleukin-2 (IL2) and interferon-gamma (IFNG). Inhibits cytokine production and T-cell effector function by repressing the activity of two key transcription factors, RELA and NFATC2 (PubMed:15790681). Mediates transcriptional repression of IL2 via its association with histone acetylase KAT5 and histone deacetylase HDAC7 (By similarity). Can activate the expression of TNFRSF18, IL2RA and CTLA4 and repress the expression of IL2 and IFNG via its association with transcription factor RUNX1 (PubMed:17377532). Inhibits the differentiation of IL17 producing helper T-cells (Th17) by antagonizing RORC function, leading to down-regulation of IL17 expression, favoring Treg development (PubMed:18368049). Inhibits the transcriptional activator activity of RORA (By similarity). Can repress the expression of IL2 and IFNG via its association with transcription factor IKZF4 (PubMed:19696312). [UniProt] |
| Highlight | Related products: FOXP3 antibodies; FOXP3 Duos / Panels; Anti-Mouse IgG secondary antibodies; Related news: Tumor-Infiltrating Lymphocytes (TILs) |
| Research Area | Cell Biology and Cellular Response antibody; Gene Regulation antibody; Immune System antibody; Regulatory T cells Study antibody |
| Calculated MW | 47 kDa |
| PTM | Polyubiquitinated, leading to its proteasomal degradation in regulatory T-cells (Treg) which is mediated by STUB1 in a HSPA1A/B-dependent manner. Deubiquitinated by USP7 leading to increase in protein stability. Phosphorylation at Ser-418 regulates its transcriptional repressor activity and consequently, regulatory T-cells (Treg) suppressive function. Dephosphorylated at Ser-418 by protein phosphatase 1 (PP1) in Treg cells derived from patients with rheumatoid arthritis. Phosphorylation by CDK2 negatively regulates its transcriptional activity and protein stability (By similarity). Acetylation on lysine residues stabilizes FOXP3 and promotes differentiation of T-cells into induced regulatory T-cells (iTregs) associated with suppressive functions. Deacetylated by SIRT1. Undergoes proteolytic cleavage in activated regulatory T-cells (Treg), and can be cleaved at either the N- or C-terminal site, or at both sites. |
Clone References
Single-cell analysis of normal and FOXP3-mutant human T cells: FOXP3 expression without regulatory T cell development.
Protein phosphatase 5 is a major component of glucocorticoid receptor.hsp90 complexes with properties of an FK506-binding immunophilin.
IP / Rabbit
Folding of the glucocorticoid receptor by the reconstituted Hsp90-based chaperone machinery. The initial hsp90.p60.hsp70-dependent step is sufficient for creating the steroid binding conformation.
IP / Mouse
Compartmentation of alpha-internexin and neurofilament triplet proteins in cultured hippocampal neurons.
WB / Rat
The basic helix-loop-helix/PAS factor Sim is associated with hsp90. Implications for regulation by interaction with partner factors.
IP / D. melanogaster
A tyrosine kinase-dependent pathway regulates ligand-dependent activation of the dioxin receptor in human keratinocytes.
IP / Human
All of the factors required for assembly of the glucocorticoid receptor into a functional heterocomplex with heat shock protein 90 are preassociated in a self-sufficient protein folding structure, a foldosome.
WB / Mouse
The 23-kDa acidic protein in reticulocyte lysate is the weakly bound component of the hsp foldosome that is required for assembly of the glucocorticoid receptor into a functional heterocomplex with hsp90.
WB / Rabbit
