FOXP3 Monoclonal Antibody (236A/E7), eBioscience™, Invitrogen™

Description

Description: The 236A/E7 antibody reacts with human Foxp3 protein also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3, a 49-55 kDa protein, is a member of the forkhead/winged-helix family of transcriptional regulators, and was identified as the gene defective in ′scurfy′ (sf) mice. Constitutive high expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg cells), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells. Intracellular staining and flow cytometric analysis of freshly isolated human peripheral blood mononuclear cells (PBMCs) with the 236A/E7 antibody using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol reveals staining of the CD4+CD25bright population. The epitope from 236A/E7 is different from that of PCH101 (cat. 72-5776). This antibody has also been shown to recognize rhesus macaque, sooty mangabey and cynomolgus macaque. Applications Tested: This 236A/E7 antibody has been tested by immunoblotting and immunohistochemistry of formalin-fixed paraffin embedded tissue using low or high pH antigen retrieval. For western blotting of reduced cell lysates (which include the nuclear fraction) the antibody can be used at 1-10 µg/mL and for IHC can be used at less than or equal to 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered. FOXP3 (Forkhead box protein 3) is a member of the forkhead/winged-helix family of transcriptional regulators, highly conserved across mammals, and essential for normal immune homeostasis. FOXP3 is 381 amino acids long, stably and constitutively expressed at a high level in CD25 + CD4 positive regulatory T cells, a low level in CD4-positive/CD25-negative cells, and is absent in CD4-negative/CD8-positive T cells. FOXP3 may be a master regulatory gene, and a more specific marker of regulatory T cells. Defects in the gene encoding FOXP3 protein cause the scurfy phenotype in mice. In humans FOXP3 defects play a role in IPEX syndrome (immune dysfunction, polyendocrinopathy, enteropathy, X-linked syndrome), also known as X-linked autoimmunity-allergic dysregulation (XLAAD) syndrome. Transcript variants of FOXP3 encoding different isoforms have been identified. In human breast and colon cancer cells, expression of FOXP3 is regulated by p53 in response to the DNA damage.