Anti-ELK1 antibody [EPR1913(2)]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ELK1 antibody [EPR1913(2)] (AB188316)

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

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ChIC/CUT&RUN sequencing - Anti-ELK1 antibody [EPR1913(2)] (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).

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-ELK1 antibody [EPR1913(2)] (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.

• WB

Lab

Western blot - Anti-ELK1 antibody [EPR1913(2)] (AB188316)

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)] (ab188316)

Predicted band size: 45 kDa

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• WB

Lab

Western blot - Anti-ELK1 antibody [EPR1913(2)] (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)] (ab188316) 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

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• WB

Supplier Data

Western blot - Anti-ELK1 antibody [EPR1913(2)] (AB188316)

All lanes:

Western blot - Anti-ELK1 antibody [EPR1913(2)] (ab188316) 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

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• WB

CiteAb

Western blot - Anti-ELK1 antibody [EPR1913(2)] (AB188316)

ELK1 western blot using anti-ELK1 antibody [EPR1913(2)] ab188316. Publication image and figure legend from Lu, W., Jia, D., et al., 2017, Sci Rep, PubMed 28827748.

ab188316 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab188316 please see the product overview.

CSEI regulated the expressions of hematopoietic function related proteins in CHRF and K562 cells. After a 24-h CSEI (0, 0.05, 0.1 mg/ml) treatment, the protein expression levels of P-RSK1p90, ELK1 and c-Myc in CHRF cells (a) and K562 cells (b), full-length blots are presented in Supplementary Fig. S5 a, and the expression levels of GATA-1 and GATA-2 in the terminal mature CHRF cells (c) and differentiated K562 cells (d) were detected by western blot, respectively, full-length blots are presented in Supplementary Fig. S5 (b). The quantitative data of these protein levels were normalized by their GAPDH expressions and expressed as a percentage of the corresponding relative intensity of control. The data are shown as the means ± S.E.M. (n = 6). *p < 0.05, **p < 0.01 and ***p < 0.001 versus 0 mg/ml CSEI-treated cells.

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