Human SREBP2 Antibody

Detection of Human SREBP2 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and Raji human Burkitt's lymphoma cell line. Gels were loaded with 15 µg of nuclear extracts (Nuc). PVDF membrane was probed with 2 µg/mL of Goat Anti-Human SREBP2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7119) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for SREBP2 at approximately 125 kDa and 60-70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of SREBP2-regulated Genes by Chromatin Immunoprecipitation. HUVEC human umbilical vein endothelial cells were serum-starved for 5 hours, fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. SREBP2/DNA complexes were immuno-precipitated using 5 µg Goat Anti-Human SREBP2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7119) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immuno-complexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. TheABCA1promoter was detected by standard PCR. In the figure, "Input" represents total cell lysate DNA.

SREBP2 in HeLa Human Cell Line. SREBP2 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human SREBP2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7119) at 15 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.

Detection of Human SREBP2 by Western Blot lincNORS regulates cholesterol synthesis via interaction with RALY.a The level of hnRNPC, RALY, TRIM28, SREBP2, and MS2-GST in the MS2 pulldown material and input lysate was detected by western blot. The data presented are a representative image from two to three independent experiments. b Subcellular location of RALY was detected by western blot. Tubulin, Lamin-B1, and acetyl-histone H3 were included as controls for cytoplasmic, nuclear, and chromatin-associated fractions. A representative image from three independent experiments is shown. c RNA immunoprecipitation (RIP) analysis of RALY. The presence of lincNORS and 18s rRNA in the precipitated complex was detected by qPCR. Data represent mean ± SD from three biological replicates (two-sided Student’s t-test). d qPCR analysis of NORS, MSMO1, and SQLE expression in MCF-7 cells transfected with control, lincNORS siRNA, and/or RALY siRNA. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test). RALY knockdown was confirmed by western blot in each experiment and a representative image is shown on the right. e Enrichment of RNA of cholesterol synthesis genes in RALY-containing immunoprecipitated complex. The barplot displays the mean fold enrichment ± S.E.M. from three independent RALY RIP-Seq vs control experiments in MCF7 cells. The statistical significance of the enrichment was determined with CuffDiff v2.2.1 (*FDR < 0.05, **FDR < 0.01, ***FDR < 0.001). Source data are provided as a Source Data file. f Enrichment of MSMO1, SQLE, H1FX, and GAPDH in RALY-containing immunoprecipitated complex after lincNORS or hnRNPC knockdown was detected by qPCR. Data represent mean ± SD from four biological replicates (two-sided Student’s t-test). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32958772), licensed under a CC-BY license. Not internally tested by R&D Systems.