Human FoxP3 Antibody

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.

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.

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. TheIL-2promoter was detected by standard PCR.