Human CHD7 Antibody

Detection of Human CHD7 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human CHD7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7350) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CHD7 at approximately 350 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

CHD7 in HepG2 Human Cell Line. CHD7 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Sheep Anti-Human CHD7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7350) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of CHD7 by Western Blot CHD7 and its ATP-dependent chromatin remodeling activities are required for sensory epithelium derivation. A Sanger sequencing chromatograms of CHD7KO/+ and CHD7KO/KO alleles cloned into TOPO vectors. b Western blotting of WT (PAX2nG), CHD7KO/+, and CHD7KO/KO hESCs using an anti-CHD7 antibody. Source data are provided as a Source Data file. c Sanger sequencing chromatograms of WT (PAX2nG), CHD7S834F/+, and CHD7S834F/S834F hESCs at the CHD7 c.2501 locus. d–w Immunostaining of d20 and d70 WT and CHD7 mutant organoids. Antibodies highlight otic progenitors (PAX2nG, PAX8, and EPCAM), hair cells (MYO7A, POU4F3, PCP4, SOX2, and F-actin for stereocilia of hair cells), and supporting cells (SPARCL1 and SOX2). Scale bars, 25 µm (top two rows of d–w), 10 µm (third row of d–w), and 5 µm (bottom row of d–w). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36396635), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CHD7 by Western Blot CHD7 is expressed at key otic development stages.a Western blotting of WT (PAX2−2a-nGFP cell line, hereafter PAX2nG) and CHD7−3×Flag hESCs using anti-Flag and anti-CHD7 antibodies. Calculated molecular weight of CHD7 and CHD7-3×Flag are 336 kDa and 339 kDa, respectively. Source data are provided as a Source Data file. b Schematics of otic lineage differentiation during human inner ear organoid culture. Schematics adapted from Nie J, Hashino E. (2020) Generation of inner ear organoids from human pluripotent stem cells. Methods in Cell Biology, 159: 303–321, with permission from Elsevier. c–g Immunostaining at key otic development stages in CHD7−3×Flag PAX2nG human inner ear organoids using an anti-Flag antibody, as well as antibodies against NNE markers TFAP2A and CDH1, OEPD markers TFAP2A and PAX8, otic placode/pit and otic vesicle markers PAX2nG, PAX8, and EPCAM, and hair cell markers MYO7A and SOX2 and supporting cell marker SOX2. Scale bars, 25 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36396635), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CHD7 by Western Blot CHD7 and its ATP-dependent chromatin remodeling activities are required for sensory epithelium derivation. A Sanger sequencing chromatograms of CHD7KO/+ and CHD7KO/KO alleles cloned into TOPO vectors. b Western blotting of WT (PAX2nG), CHD7KO/+, and CHD7KO/KO hESCs using an anti-CHD7 antibody. Source data are provided as a Source Data file. c Sanger sequencing chromatograms of WT (PAX2nG), CHD7S834F/+, and CHD7S834F/S834F hESCs at the CHD7 c.2501 locus. d–w Immunostaining of d20 and d70 WT and CHD7 mutant organoids. Antibodies highlight otic progenitors (PAX2nG, PAX8, and EPCAM), hair cells (MYO7A, POU4F3, PCP4, SOX2, and F-actin for stereocilia of hair cells), and supporting cells (SPARCL1 and SOX2). Scale bars, 25 µm (top two rows of d–w), 10 µm (third row of d–w), and 5 µm (bottom row of d–w). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36396635), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CHD7 by Western Blot CHD7 is expressed at key otic development stages.a Western blotting of WT (PAX2−2a-nGFP cell line, hereafter PAX2nG) and CHD7−3×Flag hESCs using anti-Flag and anti-CHD7 antibodies. Calculated molecular weight of CHD7 and CHD7-3×Flag are 336 kDa and 339 kDa, respectively. Source data are provided as a Source Data file. b Schematics of otic lineage differentiation during human inner ear organoid culture. Schematics adapted from Nie J, Hashino E. (2020) Generation of inner ear organoids from human pluripotent stem cells. Methods in Cell Biology, 159: 303–321, with permission from Elsevier. c–g Immunostaining at key otic development stages in CHD7−3×Flag PAX2nG human inner ear organoids using an anti-Flag antibody, as well as antibodies against NNE markers TFAP2A and CDH1, OEPD markers TFAP2A and PAX8, otic placode/pit and otic vesicle markers PAX2nG, PAX8, and EPCAM, and hair cell markers MYO7A and SOX2 and supporting cell marker SOX2. Scale bars, 25 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36396635), licensed under a CC-BY license. Not internally tested by R&D Systems.