|Detection of Human HNF‑3 beta /FoxA2 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, Jurkat human acute T cell leukemia cell line, and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human HNF‑3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for HNF‑3 beta /FoxA2 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Chromatin Immunoprecipitation (ChIP)
|Detection of HNF‑3 beta /FoxA2-regulated Genes by Chromatin Immunoprecipitation. Mouse splenocytes were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HNF‑3 beta /FoxA2/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human HNF‑3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) 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 E-RABP promoter was detected by standard PCR.|
|HNF‑3 beta /FoxA2 in Endoderm Differentiated BG01V Human Stem Cells. HNF‑3 beta /FoxA2 was detected in immersion fixed endoderm differentiated BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human HNF‑3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
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