Anti-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade - BSA and Azide free

• WB

Lab

Western blot - Anti-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade - BSA and Azide free (AB248517)

This data was developed using the same antibody clone in a different buffer formulation (ab133482).

Lanes 1-4 : Merged signal (red and green). Green - ab133482 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133482 Anti-EHMT2/G9A antibody [EPR4019(2)] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab133482 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade (<a href='/en-us/products/primary-antibodies/ehmt2-g9a-antibody-epr40192-chip-grade-ab133482'>ab133482</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

EHMT2/G9A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human EHMT2 (G9A) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ehmt2-g9a-knockout-hela-cell-line-ab265149'>ab265149</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 132 kDa

Observed band size: 160 kDa

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• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade - BSA and Azide free (AB248517)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133482)

CUT&RUN profiling with EHMT2 antibody reveals the expected genomic enrichment pattern in cells. Representative genome browser tracks show CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with EHMT2 antibody (Abcam ab133482, 0.5 µg). 500,000 HeLa cells were used per reaction. IgG, H3K4me3, and H3K27me3 antibodies were included as controls to assess non-specific background, active promoters, and repressed chromatin, respectively. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Images were generated using Integrative Genomics Viewer (IGV, Broad Institute).

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade - BSA and Azide free (AB248517)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133482)

CUT&RUN profiling with EHMT2 antibody demonstrates robust genome-wide enrichment in cells. Heatmaps of genome-wide signal flanking annotated transcription start sites (TSSs, +/- 2 kbp) display CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with EHMT2 antibody (Abcam ab133482, 0.5 µg). 500,000 HeLa cells were used per reaction. IgG antibody was included as a negative control to assess non-specific background. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Heatmaps were generated using ChAsE (Younesy et al., Bioinformatics 2016; PMID 27378294). Row-linked data are ranked by intensity relative to EHMT2, with red indicating high localized enrichment and blue denoting background.