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Bethyl anti brd4
Anti Brd4, supplied by Bethyl, used in various techniques. Bioz Stars score: 96/100, based on 513 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 513 article reviews
anti brd4 - by Bioz Stars, 2026-10
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Article Title: Androgen receptor–mediated assisted loading of the glucocorticoid receptor modulates transcriptional responses in prostate cancer cells
Article Snippet: Primary antibodies were as follows: anti-GR (Santa Cruz Biotechnology sc-1003, RRID:AB_631572), anti-GR (Cell Signaling Technology 12041, RRID:AB_2631286), anti-AR , anti-FOXA1 (Abcam ab23738, RRID:AB_2104842), anti-SMARCA4 (Abcam ab110641, RRID:AB_10861578), anti-BRD4 (Bethyl Laboratories A301-985A, RRID:AB_1576498), anti-GAPDH (Santa Cruz Biotechnology sc-25778, RRID: AB_10167668), antitubulin (Santa Cruz Biotechnology sc-5286, RRID:AB_628411), anti-BrdU (Abcam ab6326, RRID:AB_305426), and normal rabbit IgG (Millipore 12-370, RRID:AB_145841).

Isolation:

Article Title: Regulation of inflammatory responses by pH-dependent transcriptional condensates.
Article Snippet: .. The isolated nuclei were washed (10 mM Tris-HCl pH 8, 1 mM EDTA, 0.5 mM EGTA, and 200 mM NaCl) and sheared in (0.1% sodium dodecyl sulfate (SDS), 1 mM EDTA, and 10 mM Tris-HCl pH 8) with the Covaris E229 sonicator for 10 min. After centrifugation, chromatin was immunoprecipitated overnight at 4 ◦ C with 1:100 anti-BRD9 (Bethyl, pAb #A700-153) or 5 μg anti-BRD4 (Bethyl, Rabbit pAb #A301-985A50).The next day, the antibody-bound DNA was ll OPEN ACCESS Cell 188, 5632–5652.e1–e13, October 2, 2025 e7 Article incubated with Protein A+G Dynabeads (Invitrogen) in ChIP buffer (50 mM Hepes-KOH pH 7.5, 300 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate), washed, and treated with Proteinase K and RNase A and reverse cross linked. .. Purified ChIP DNA was used for library generation (NEBNext Ultra II DNA Library Prep Kit for Illumina) according to manufacturer’s instructions.

Centrifugation:

Article Title: Regulation of inflammatory responses by pH-dependent transcriptional condensates.
Article Snippet: .. The isolated nuclei were washed (10 mM Tris-HCl pH 8, 1 mM EDTA, 0.5 mM EGTA, and 200 mM NaCl) and sheared in (0.1% sodium dodecyl sulfate (SDS), 1 mM EDTA, and 10 mM Tris-HCl pH 8) with the Covaris E229 sonicator for 10 min. After centrifugation, chromatin was immunoprecipitated overnight at 4 ◦ C with 1:100 anti-BRD9 (Bethyl, pAb #A700-153) or 5 μg anti-BRD4 (Bethyl, Rabbit pAb #A301-985A50).The next day, the antibody-bound DNA was ll OPEN ACCESS Cell 188, 5632–5652.e1–e13, October 2, 2025 e7 Article incubated with Protein A+G Dynabeads (Invitrogen) in ChIP buffer (50 mM Hepes-KOH pH 7.5, 300 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate), washed, and treated with Proteinase K and RNase A and reverse cross linked. .. Purified ChIP DNA was used for library generation (NEBNext Ultra II DNA Library Prep Kit for Illumina) according to manufacturer’s instructions.

Immunoprecipitation:

Article Title: Regulation of inflammatory responses by pH-dependent transcriptional condensates.
Article Snippet: .. The isolated nuclei were washed (10 mM Tris-HCl pH 8, 1 mM EDTA, 0.5 mM EGTA, and 200 mM NaCl) and sheared in (0.1% sodium dodecyl sulfate (SDS), 1 mM EDTA, and 10 mM Tris-HCl pH 8) with the Covaris E229 sonicator for 10 min. After centrifugation, chromatin was immunoprecipitated overnight at 4 ◦ C with 1:100 anti-BRD9 (Bethyl, pAb #A700-153) or 5 μg anti-BRD4 (Bethyl, Rabbit pAb #A301-985A50).The next day, the antibody-bound DNA was ll OPEN ACCESS Cell 188, 5632–5652.e1–e13, October 2, 2025 e7 Article incubated with Protein A+G Dynabeads (Invitrogen) in ChIP buffer (50 mM Hepes-KOH pH 7.5, 300 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate), washed, and treated with Proteinase K and RNase A and reverse cross linked. .. Purified ChIP DNA was used for library generation (NEBNext Ultra II DNA Library Prep Kit for Illumina) according to manufacturer’s instructions.

Article Title: BRD4 acts as a transcriptional repressor of RhoB to inhibit terminal erythropoiesis.
Article Snippet: .. The following primary antibodies were used for immunoprecipitation or Western blotting: anti-IgG (Abcam, ab172730), anti-BRD4 (Bethyl, A301-985 A, Abcam, ab243862), antiActin (ABclonal, AC038), anti-EHMT1 (CST, 35005s), and anti-RhoB (CST, 2098T). ..

Incubation:

Article Title: Regulation of inflammatory responses by pH-dependent transcriptional condensates.
Article Snippet: .. The isolated nuclei were washed (10 mM Tris-HCl pH 8, 1 mM EDTA, 0.5 mM EGTA, and 200 mM NaCl) and sheared in (0.1% sodium dodecyl sulfate (SDS), 1 mM EDTA, and 10 mM Tris-HCl pH 8) with the Covaris E229 sonicator for 10 min. After centrifugation, chromatin was immunoprecipitated overnight at 4 ◦ C with 1:100 anti-BRD9 (Bethyl, pAb #A700-153) or 5 μg anti-BRD4 (Bethyl, Rabbit pAb #A301-985A50).The next day, the antibody-bound DNA was ll OPEN ACCESS Cell 188, 5632–5652.e1–e13, October 2, 2025 e7 Article incubated with Protein A+G Dynabeads (Invitrogen) in ChIP buffer (50 mM Hepes-KOH pH 7.5, 300 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate), washed, and treated with Proteinase K and RNase A and reverse cross linked. .. Purified ChIP DNA was used for library generation (NEBNext Ultra II DNA Library Prep Kit for Illumina) according to manufacturer’s instructions.

Article Title: BRD4 acts as a transcriptional repressor of RhoB to inhibit terminal erythropoiesis.
Article Snippet: 75,000 erythroblasts treated with DMSO or MS436 for 2 days were prepared for each sample, and technical triplicate were performed with Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903) following the manufacturer’s instructions. .. Briefly, erythroblasts were incubated with Concanavalin A beads and anti-BRD4 (Bethyl, A301-985 A) antibody overnight at 4 °C. ..

Chromatin Immunoprecipitation:

Article Title: Regulation of inflammatory responses by pH-dependent transcriptional condensates.
Article Snippet: .. The isolated nuclei were washed (10 mM Tris-HCl pH 8, 1 mM EDTA, 0.5 mM EGTA, and 200 mM NaCl) and sheared in (0.1% sodium dodecyl sulfate (SDS), 1 mM EDTA, and 10 mM Tris-HCl pH 8) with the Covaris E229 sonicator for 10 min. After centrifugation, chromatin was immunoprecipitated overnight at 4 ◦ C with 1:100 anti-BRD9 (Bethyl, pAb #A700-153) or 5 μg anti-BRD4 (Bethyl, Rabbit pAb #A301-985A50).The next day, the antibody-bound DNA was ll OPEN ACCESS Cell 188, 5632–5652.e1–e13, October 2, 2025 e7 Article incubated with Protein A+G Dynabeads (Invitrogen) in ChIP buffer (50 mM Hepes-KOH pH 7.5, 300 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate), washed, and treated with Proteinase K and RNase A and reverse cross linked. .. Purified ChIP DNA was used for library generation (NEBNext Ultra II DNA Library Prep Kit for Illumina) according to manufacturer’s instructions.

Article Title: Identification of super-enhancer-based biomarkers for predicting survival and immunotherapy efficacy in colorectal cancer
Article Snippet: .. ChIP assays were conducted as reported in previous studies with the anti-H3K27ac (Abcam, ab177178, Cambridge, UK), anti-P300 (Abcam, ab275378, Cambridge, UK) and anti-BRD4 (Bethyl Laboratories, A301-985A100, Montgomery, TX, USA) antibodies. ..

Western Blot:

Article Title: BRD4 acts as a transcriptional repressor of RhoB to inhibit terminal erythropoiesis.
Article Snippet: .. The following primary antibodies were used for immunoprecipitation or Western blotting: anti-IgG (Abcam, ab172730), anti-BRD4 (Bethyl, A301-985 A, Abcam, ab243862), antiActin (ABclonal, AC038), anti-EHMT1 (CST, 35005s), and anti-RhoB (CST, 2098T). ..



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BHB-mediated HDAC inhibition shifts hepatic anabolism toward lipid oxidation. (A) Experimental design. (B) Serum gross appearance and the FFA levels ( n = 5). (C) Body weight loss of mice with the indicated treatment ( n = 5). (D and E) ChIP-seq data displaying H3K27ac (D) or <t>BRD4</t> (E) occupancy at two subsets of genes: H3K27ac and BRD4 enriched at the genomic loci of fasting-induced transcripts (energy mobilization associated genes, such as Cyp4a14), but dislodged from those loci of fasting-suppressed transcripts (energy storage-associated genes, such as Pcsk9). (ref. to Hsieh et al., 2022, Mol Cell for detail method of analysis). (F and G) GO enrichment analysis of the fasting-induced H3K27ac (F) and BRD4 (G) peak-associated genes. The reads number of the ChIP-seq data for H3K27ac (log2FC > 1 and P < 0.01) or BRD4 (log2FC > 1.2 and P < 0.01) were included as the fasting-induced peaks. (H and I) Gene tracks display that fasting induced the occupancy of H3K27ac and BRD4 at the genomic loci of energy mobilization-associated genes (Cyp4a14, Pck1) (H), but reduced at those loci of energy storage-associated genes (Pcsk9, Srebf1) (I). (J) The ratio of the liver to body weight (upper panel) and the liver triglycerides levels (lower panel) of mice with the indicated treatment ( n = 5). (K-O) Body weight loss data (K), serum gross appearance and FFA levels (L), the blood glucose and BHB levels (M), the ratio of liver to body weight (N) and Liver triglycerides (O)of Brd 4 -flox or Brd4 hKO mice subjected to 24 h of fasting ( n = 5). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (K-O) or one-way ANOVA followed with Bonferroni’s multiple comparisons test (B, C, and J).
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Effect of AZD5153 on protein expression in irradiated pancreatic cancer cells, n=6. ( a and b ) Western blot analysis was performed to detect the expression of <t>BRD4,</t> c-Myc in different groups, β-actin was used as a loading control. ( c and d ) The protein expression of p-ATM, ATM, p-chk1, chk1, p-cdc25, cdc25, p-cdc2, cdc2 in different groups was detected by western blot, β-actin was used as a loading control. Results shown are the means ± SD of 3 independent experiments with similar results for all assays. Significance was determined by Student’s t -test (* p < 0.05, ** p < 0.01, *** p < 0.001).
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Image Search Results


BHB-mediated HDAC inhibition shifts hepatic anabolism toward lipid oxidation. (A) Experimental design. (B) Serum gross appearance and the FFA levels ( n = 5). (C) Body weight loss of mice with the indicated treatment ( n = 5). (D and E) ChIP-seq data displaying H3K27ac (D) or BRD4 (E) occupancy at two subsets of genes: H3K27ac and BRD4 enriched at the genomic loci of fasting-induced transcripts (energy mobilization associated genes, such as Cyp4a14), but dislodged from those loci of fasting-suppressed transcripts (energy storage-associated genes, such as Pcsk9). (ref. to Hsieh et al., 2022, Mol Cell for detail method of analysis). (F and G) GO enrichment analysis of the fasting-induced H3K27ac (F) and BRD4 (G) peak-associated genes. The reads number of the ChIP-seq data for H3K27ac (log2FC > 1 and P < 0.01) or BRD4 (log2FC > 1.2 and P < 0.01) were included as the fasting-induced peaks. (H and I) Gene tracks display that fasting induced the occupancy of H3K27ac and BRD4 at the genomic loci of energy mobilization-associated genes (Cyp4a14, Pck1) (H), but reduced at those loci of energy storage-associated genes (Pcsk9, Srebf1) (I). (J) The ratio of the liver to body weight (upper panel) and the liver triglycerides levels (lower panel) of mice with the indicated treatment ( n = 5). (K-O) Body weight loss data (K), serum gross appearance and FFA levels (L), the blood glucose and BHB levels (M), the ratio of liver to body weight (N) and Liver triglycerides (O)of Brd 4 -flox or Brd4 hKO mice subjected to 24 h of fasting ( n = 5). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (K-O) or one-way ANOVA followed with Bonferroni’s multiple comparisons test (B, C, and J).

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: BHB-mediated HDAC inhibition shifts hepatic anabolism toward lipid oxidation. (A) Experimental design. (B) Serum gross appearance and the FFA levels ( n = 5). (C) Body weight loss of mice with the indicated treatment ( n = 5). (D and E) ChIP-seq data displaying H3K27ac (D) or BRD4 (E) occupancy at two subsets of genes: H3K27ac and BRD4 enriched at the genomic loci of fasting-induced transcripts (energy mobilization associated genes, such as Cyp4a14), but dislodged from those loci of fasting-suppressed transcripts (energy storage-associated genes, such as Pcsk9). (ref. to Hsieh et al., 2022, Mol Cell for detail method of analysis). (F and G) GO enrichment analysis of the fasting-induced H3K27ac (F) and BRD4 (G) peak-associated genes. The reads number of the ChIP-seq data for H3K27ac (log2FC > 1 and P < 0.01) or BRD4 (log2FC > 1.2 and P < 0.01) were included as the fasting-induced peaks. (H and I) Gene tracks display that fasting induced the occupancy of H3K27ac and BRD4 at the genomic loci of energy mobilization-associated genes (Cyp4a14, Pck1) (H), but reduced at those loci of energy storage-associated genes (Pcsk9, Srebf1) (I). (J) The ratio of the liver to body weight (upper panel) and the liver triglycerides levels (lower panel) of mice with the indicated treatment ( n = 5). (K-O) Body weight loss data (K), serum gross appearance and FFA levels (L), the blood glucose and BHB levels (M), the ratio of liver to body weight (N) and Liver triglycerides (O)of Brd 4 -flox or Brd4 hKO mice subjected to 24 h of fasting ( n = 5). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (K-O) or one-way ANOVA followed with Bonferroni’s multiple comparisons test (B, C, and J).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: Inhibition, ChIP-sequencing, Two Tailed Test

Glucose-mediated histone acetylation promotes lipogenesis. (A-D) ChIP-seq data showed that re-feeding induced a significant redistribution of H3K27ac and BRD4 in the whole genome (A and B) with an increased occupancy at the genomic loci of lipids synthesis genes ( Srebf1 , Pcsk9 ) (C) but a decreased occupancy at those of fatty acids catabolic ( Cyp4a14 ) and gluconeogenic genes ( Pck1 ) (D). (E and F) qPCR assay showed that re-feeding induced the expression of glucose utilization ( Gck ) and lipid synthesis genes ( Pcsk9 , Hmgcr , Dgat1 , Srebf1 ) in the liver (E), while the expression was suppressed by JQ35 treatment (F) ( n = 4). (G and H) Brd 4 -flox or Brd 4 -hKO mice housed in a thermoneutral environment (30°C) feeding with high-fat diet. Hepatic Brd4 knockout suppressed body weight gain (G) without influence food intake (H) ( n = 5–6). (I-L) ITT assays (I) , liver weight (J), the liver gross appearance, HE and Oil Red O staining (K), and TG levels (L) of Brd 4 -flox or Brd 4 -hKO mice that were subjected to HFD feeding with housing at 30°C ( n = 5–6). (M) qPCR assay of lipids anabolism- ( Cd36 , Pparg , Dgat1 ) and VLDLs secretion/metabolism- ( Mttp , Apoc3 ) associated genes in the liver of Brd 4 -flox or Brd 4 -hKO mice ( n = 5–6). (N) The serum ALT levels were lower in Brd 4 -hKO than that of control mice ( n = 5–6). (O-Q) WAT weight (O) and serum TG (P) and FFA (Q) levels of Brd 4 -flox or Brd 4 -hKO mice under HFD feeding ( n = 5–6). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (E, F, H, J, and L-Q) or two-way ANOVA followed with Bonferroni’s multiple comparisons test (G and I).

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: Glucose-mediated histone acetylation promotes lipogenesis. (A-D) ChIP-seq data showed that re-feeding induced a significant redistribution of H3K27ac and BRD4 in the whole genome (A and B) with an increased occupancy at the genomic loci of lipids synthesis genes ( Srebf1 , Pcsk9 ) (C) but a decreased occupancy at those of fatty acids catabolic ( Cyp4a14 ) and gluconeogenic genes ( Pck1 ) (D). (E and F) qPCR assay showed that re-feeding induced the expression of glucose utilization ( Gck ) and lipid synthesis genes ( Pcsk9 , Hmgcr , Dgat1 , Srebf1 ) in the liver (E), while the expression was suppressed by JQ35 treatment (F) ( n = 4). (G and H) Brd 4 -flox or Brd 4 -hKO mice housed in a thermoneutral environment (30°C) feeding with high-fat diet. Hepatic Brd4 knockout suppressed body weight gain (G) without influence food intake (H) ( n = 5–6). (I-L) ITT assays (I) , liver weight (J), the liver gross appearance, HE and Oil Red O staining (K), and TG levels (L) of Brd 4 -flox or Brd 4 -hKO mice that were subjected to HFD feeding with housing at 30°C ( n = 5–6). (M) qPCR assay of lipids anabolism- ( Cd36 , Pparg , Dgat1 ) and VLDLs secretion/metabolism- ( Mttp , Apoc3 ) associated genes in the liver of Brd 4 -flox or Brd 4 -hKO mice ( n = 5–6). (N) The serum ALT levels were lower in Brd 4 -hKO than that of control mice ( n = 5–6). (O-Q) WAT weight (O) and serum TG (P) and FFA (Q) levels of Brd 4 -flox or Brd 4 -hKO mice under HFD feeding ( n = 5–6). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (E, F, H, J, and L-Q) or two-way ANOVA followed with Bonferroni’s multiple comparisons test (G and I).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: ChIP-sequencing, Expressing, Knock-Out, Staining, Control, Two Tailed Test

Effect of AZD5153 on protein expression in irradiated pancreatic cancer cells, n=6. ( a and b ) Western blot analysis was performed to detect the expression of BRD4, c-Myc in different groups, β-actin was used as a loading control. ( c and d ) The protein expression of p-ATM, ATM, p-chk1, chk1, p-cdc25, cdc25, p-cdc2, cdc2 in different groups was detected by western blot, β-actin was used as a loading control. Results shown are the means ± SD of 3 independent experiments with similar results for all assays. Significance was determined by Student’s t -test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Journal: Drug Design, Development and Therapy

Article Title: The Effect of AZD5153 on Radiosensitivity in Pancreatic Cancer Cells Through ATM-chk1 Pathway

doi: 10.2147/DDDT.S568551

Figure Lengend Snippet: Effect of AZD5153 on protein expression in irradiated pancreatic cancer cells, n=6. ( a and b ) Western blot analysis was performed to detect the expression of BRD4, c-Myc in different groups, β-actin was used as a loading control. ( c and d ) The protein expression of p-ATM, ATM, p-chk1, chk1, p-cdc25, cdc25, p-cdc2, cdc2 in different groups was detected by western blot, β-actin was used as a loading control. Results shown are the means ± SD of 3 independent experiments with similar results for all assays. Significance was determined by Student’s t -test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The membranes were blocked with 5% skim milk for 2 h, and incubated with primary antibodies against BRD4 (Cell Signaling Technology, Cat# 13440s), c-Myc (Cell Signaling Technology, Cat# 18583s), p-ATM (Abcam, Cat# ab81292), ATM (Cell Signaling Technology, Cat# 2873s), p-cdc25C (Cell Signaling Technology, Cat# 4901s), cdc25C (Cell Signaling Technology, Cat# 4688s), p-chk1 (Cell Signaling Technology, Cat# 2348s), chk1 (Abcam, Cat# ab40866), p-cdc2 (Cell Signaling Technology, Cat# 4539s), cdc2 (Cell Signaling Technology, Cat# 9116s), γ-H2AX (Cell Signaling Technology, Cat# 9718s), cleaved PARP (Cell Signaling Technology, Cat# 5625s), Bax (Cell Signaling Technology, Cat# 14796s), β-actin (Abcam, Cat# ab6276),Vinculin (Cell Signaling Technology, Cat# 13901s) overnight at 4 °C.

Techniques: Expressing, Irradiation, Western Blot, Control