Human KLF4 Antibody

Detection of Human KLF4 by Western Blot. Western blot shows lysates of HT-29 human colon adenocarcinoma cell line, SW480 human colorectal adenocarcinoma cell line, and HCT-116 human colorectal carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for KLF4 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 5.

KLF4 in BG01V Human Stem Cells. KLF4 was detected in immersion fixed BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of KLF4-regulated Genes by Chromatin Immunoprecipitation. BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. KLF4/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) or control antibody (AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (MAG999) and DNA was purified using chelating resin solution. TheB2Rpromoter was detected by standard PCR.

KLF4 in Human Colon. KLF4 was detected in immersion fixed paraffin-embedded sections of human colon using 15 µg/mL Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) 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). Specific labeling was localized to the nuclei of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human KLF4 by Simple Western^TM. Simple Western lane view shows lysates of HT-29 human colon adenocarcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for KLF4 at approximately 63 kDa (as indicated) using 25 µg/mL of Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Detection of KLF4 in HT‑29 Human Colon Adenocarcinoma Cell Line (positive) and HDLM‑2 Human Hodgkin’s Lymphoma Cell Line (negative) Cells. KLF4 was detected in immersion fixed HT‑29 Human Colon Adenocarcinoma Cell Line (positive) and HDLM‑2 Human Hodgkin’s Lymphoma Cell Line (negative) Cells using Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.