Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free (AB250948)

This data was developed using ab188316, the same antibody clone in a different buffer formulation.

Immunocytochemical analysis of HeLa cells fixed in 2% paraformaldehyde labeling ELK1 with ab188316 at 1/100 dilution and Goat anti rabbit IgG(Alexa Fluor® 488) at 1/200 dilution. Counterstained with DAPI (blue).

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free (AB250948)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188316)

CUT&RUN profiling with ELK1 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 ELK1 antibody (Abcam ab188316, 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 ELK1, with red indicating high localized enrichment and blue denoting background.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free (AB250948)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188316)

CUT&RUN profiling with ELK1 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 ELK1 antibody (Abcam ab188316, 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).

• WB

Supplier Data

Western blot - Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free (AB250948)

This data was developed using ab188316, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ELK1 antibody [EPR1913(2)] (<a href='/en-us/products/primary-antibodies/elk1-antibody-epr19132-ab188316'>ab188316</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

K562 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 45 kDa

false

• WB

Lab

Western blot - Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free (AB250948)

This data was developed using ab188316, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)

Lane 2 : ELK1 knockout HAP1 whole cell lysate (20 μg)

Lane 3 : HeLa positve whole cell lysate (20 μg)

Lane 4 : MCF7 negative whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab188316 observed at 55 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab188316 was shown to specifically react with ELK1 in wild-type HAP1 cells as signal was lost in ELK1 knockout cells. Wild-type and ELK1 knockout samples were subjected to SDS-PAGE. ab188316 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ELK1 antibody [EPR1913(2)] (<a href='/en-us/products/primary-antibodies/elk1-antibody-epr19132-ab188316'>ab188316</a>)

Predicted band size: 45 kDa

false

• WB

Lab

Western blot - Anti-ELK1 antibody [EPR1913(2)] - BSA and Azide free (AB250948)

This data was developed using the same antibody clone in a different buffer formulation (ab188316).

Lanes 1- 2 : Merged signal (red and green). Green - ab188316 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab188316 was shown to react with ELK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261764 (knockout cell lysate ab256904) was used. Wild-type HeLa and ELK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab188316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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-ELK1 antibody [EPR1913(2)] (<a href='/en-us/products/primary-antibodies/elk1-antibody-epr19132-ab188316'>ab188316</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ELK1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ELK1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-elk1-knockout-hela-cell-line-ab261764'>ab261764</a>)

Predicted band size: 45 kDa

Observed band size: 55 kDa

false