Anti-cbx7 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-cbx7 antibody (AB21873)

IHC image of cbx7 staining in Human normal prostate formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab21873, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Lab

Western blot - Anti-cbx7 antibody (AB21873)

All lanes:

Western blot - Anti-cbx7 antibody (ab21873) at 1 µg

Lane 1:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

Lane 2:

Jurkat nuclear extract lysate (<a href='/en-us/products/unavailable/jurkat-nuclear-extract-lysate-ab14844'>ab14844</a>) at 10 µg

Lane 3:

Western blot - Hep G2 nuclear extract lysate (<a href='/en-us/products/cell-lysates/hep-g2-nuclear-extract-lysate-ab14660'>ab14660</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 28 kDa

Observed band size: 100 kDa,34 kDa,70 kDa

true

Exposure time: 30s

• WB

Project

Western blot - Anti-cbx7 antibody (AB21873)

All lanes:

Western blot - Anti-cbx7 antibody (ab21873) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

Lane 3:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 4:

Jurkat nuclear extract lysate (<a href='/en-us/products/unavailable/jurkat-nuclear-extract-lysate-ab14844'>ab14844</a>) at 10 µg

Lane 5:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 6:

Western blot - Hep G2 nuclear extract lysate (<a href='/en-us/products/cell-lysates/hep-g2-nuclear-extract-lysate-ab14660'>ab14660</a>) at 10 µg

Secondary

All lanes:

Goat Polyclonal to rabbit IgG - H&L - Pre adsorbed (HRP) at 1/3000 dilution

Predicted band size: 28 kDa

Observed band size: 100 kDa,34 kDa,70 kDa

false

• WB

CiteAb

Western blot - Anti-cbx7 antibody (AB21873)

Western Blotting using Anti-cbx7 antibody, ab21873. Publication image from Hu, S. et al., 2020, Genome Biol, 32093739. Legend direct from paper.

Non-classical function of CBX7 for the maintenance of pluripotency. a Heatmap showing CBX7, CBX7’s classical substrates H3K27me3 and H3K9me3, predicted cofactor NANOG, and H3K27ac enrichment around CBX7 ChIP-seq peak centers. Rows represent CBX7 binding sites and are ranked by normalized H3K27me3 signals. The colors indicate the normalized ChIP-seq enrichment level and the values are scaled by row. CBX7, NANOG, H3K27me3, H3K9me3, and H3K27ac ChIP-seq data were obtained from GSE64008, GSE90893, GSE58023, GSE90895, and GSE67867. b Stacked bar plot showing the percentages of classical sites, non-classical sites, and the whole genome that reside in promoter, gene body, and intergenic regions. The promoter is defined as ± 3 kb around TSS of the gene. c Real-time qPCR analysis for the expression of CBX7 and NANOG in wild type, Cbx7-knockout, and Cbx7-knockdown mESCs. Error bars represent the standard deviation for triplicate experiments, and unpaired t test with Welch’s correction was used to calculate the statistical significance for comparison (*p value < 0.05, **p value < 0.01, ***p value < 0.001, and n.s represents non-significant). d Western blot analysis of CBX7 and NANOG level in wild type, Cbx7-knockout, and Cbx7-knockdown mESCs. Tubulin was used as a loading control. Blots were cut before antibody application. Gel images for Western blot are shown in Additional file 1 : Fig. S6a. e The UCSC genome browser view of CBX7, H3K27me3, H3K9me3, NANOG, and H3K27ac enrichment at a previously reported enhancer of Nanog [37]. Enhancer loci are shaded in purple and signals represent ChIP-seq RPM. f Immunofluorescence staining for NANOG (red), OCT4 (green), DAPI (blue), and merged images in wild type, Cbx7-knockout, and Cbx7-knockdown mESCs. g The fluorescence intensity of NANOG for about 300 nuclei in wild type, Cbx7-knockout, and Cbx7-knockdown mESCs. Unpaired t test with Welch’s correction was used to calculate statistical significance for comparison

false