Detection of Human FoxP3 by Western Blot. Western blot shows lysates of human CD4+CD25+ T cells and human CD4+CD25- T cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human FoxP3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3240) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for FoxP3 at approximately 47 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of FoxP3-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. FoxP3/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human FoxP3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3240) or control antibody (Catalog # AB‑108‑C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The IL-2 promoter was detected by standard PCR.
FoxP3 in Human Tonsil. FoxP3 was detected in immersion fixed paraffin-embedded sections of human tonsil using 10 µg/mL Goat Anti-Human FoxP3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3240) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Human FoxP3 is a 47 kDa member of the P subclass of the FOX (forkhead box) family of transcription factors. It contains a Leu-rich repeat, a C2H2 zinc finger region, and a C-terminal FKH (fork head), DNA-binding domain. Three isoforms for FoxP3 have been reported. All three isoforms share the sequence used as the immunogen. FoxP3 directly associates with NFAT and NFkB, suppressing their activity in CD4+ T cells. In human, FoxP3 is found in CD4+, CD8+ and CD4+CD25+ T cells. Over the region used for immunization of the amino acid sequence, mouse FoxP3 is 83% to 88% identical to rat, human, canine, and bovine FoxP3.
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