Description: The 236A/E7 antibody reacts with human foxp3 protein also known as FORKHEAD BOX P3, SCURFIN, and JM2; cross reactivity of this antibody to other proteins has not been determined. 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 Staining Buffer Set (cat. 00-5523) and protocol reveals staining of the CD4+CD25^bright population.
The epitope from 236A/E7 is different from that of PCH101 (cat. 72-5776).
Applications Reported: This 236A/E7 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 236A/E7 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins" located at www.thermofisher.com/flowprotocols. This may be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 667 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: Forkhead box protein P3; Forkhead box protein P3 41 kDa form; Forkhead box protein P3, C-terminally processed; FOXP3delta7; immune dysregulation, polyendocrinopathy, enteropathy, X-linked; immunodeficiency, polyendocrinopathy, enteropathy, X-linked; MGC141961; MGC141963; regulatory protein Foxp3; RP23-54C14.1; Scurfin
Gene Aliases: AIID; DIETER; FOXP3; IPEX; JM2; PIDX; XPID
UniProt ID: (Human) Q9BZS1