BRD4 CUTANA™ CUT&RUN Antibody

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Figure 2: BRD4 CUT&RUN peak enrichment and functional overlap
The CUT&RUN data from Figure 1 was subjected to peak calling using MACS2. BRD4 peaks overlapped with BRG1/ SMARCA4 antibody CUT&RUN peaks (EpiCypher 13-2002, top), as has been demonstrated in the literature (Conrad et al., 2017). Three representative loci show BRD4 peaks in relation to control and BRG1 antibodies (bottom, Integrative Genomics Viewer).

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Figure 3: Western blot detection of human BRD4
Whole cell lysates were isolated from HeLa, HEK293T (“293T”), Jurkat, MB231, Hep-G2, A-549, SW620, SK-MEL-28 (“SK”), and Ramos cells using NETN lysis buffer. Lysates (15 µg) were loaded onto 4-8% SDS-PAGE gel and analyzed under standard western blot conditions using BRD4 antibody (1:1,000 dilution).

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Figure 4: Western blot detection of mouse BRD4
Whole cell lysates were isolated from NIH 3T3 (“3T3”) cells using NETN lysis buffer. Lysates (15 µg) were loaded onto 4-8% SDS-PAGE gel and analyzed under standard western blot conditions using BRD4 antibody (0.1 µg/mL).

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Figure 5: Immunoprecipitation of human BRD4
EpiCypher BRD4 antibody (3 µg) was used to immunoprecipitate whole cell lysates isolated from HeLa cells using NETN lysis buffer (1 mg per IP). A negative control IgG antibody and positive control antibodies to a different BRD4 epitopes (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto 4-8% SDS-PAGE gel (20% of IP loaded) and probed via western blot with a different BRD4 antibody (Bethyl Laboratories, Catalog No. A700-004, 1:1,000).

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Figure 6: Immunohistochemistry detection of human and mouse BRD4
(A) FFPE section of human ovarian carcinoma examined using BRD4 antibody (1:5,000 dilution, 0.2 µg/mL). (B) FFPE section of mouse CT26 colon carcinoma examined using BRD4 antibody (1:1,000 dilution, 1 µg/mL).