BRD4 Antibody (YA3486)

BRD4 Antibody (YA3486) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to BRD4.

BRD4 protein functions as a chromatin reader, specifically recognizing and binding acetylated histones, thereby playing a crucial role in transmitting epigenetic memory across cell divisions and regulating transcription. It remains associated with acetylated chromatin throughout the cell cycle, preserving the acetylated chromatin status and maintaining high-order chromatin structure, contributing to epigenetic memory for postmitotic G1 gene transcription. During interphase, BRD4 is pivotal in regulating the transcription of signal-inducible genes by associating with the P-TEFb complex and recruiting it to promoters. Additionally, it collaborates with JMJD6 in recruiting the P-TEFb complex to distal enhancers, known as anti-pause enhancers, displacing negative regulators and transforming P-TEFb into an active form that can phosphorylate the C-terminal domain of RNA polymerase II. BRD4 also acts as an atypical protein kinase, promoting phosphorylation of 'Ser-2' of the C-terminal domain of RNA polymerase II. Beyond histones, BRD4 recognizes and binds acetylated RELA, facilitating the recruitment of the P-TEFb complex and activating NF-kappa-B. Furthermore, it acts as a regulator of p53/TP53-mediated transcription, being recruited to specific target promoters upon phosphorylation by CK2. In the DNA damage response pathway, BRD4 acts as a chromatin insulator, inhibiting DNA damage response signaling by recruiting the condensin-2 complex to acetylated histones and limiting the spreading of histone H2AX/H2A.x phosphorylation. These diverse functions underscore the versatile role of BRD4 in epigenetic regulation, transcriptional control, and maintenance of chromatin structure.

Virus

O60885

Predicted band size: 152 kDa; Observed band size: 152 kDa

Protein A affinity purified.

Non-conjugated

Unmodified

AB_3102781

Epigenetics and Nuclear Signaling

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:1000; IHC: 1:50-1:100; IF: 1:50-1:200; IP: 1:50

• 
  Western blot analysis of extracts from Hela (lane 2(20μg), HepG2 (lane 3(20μg) and RAW264.7(lane 2(20μg) using BRD4 (HY-P80976) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of HepG2 cells labeling BRD4 with BRD4 antibody (HY-P80976) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with BRD4 antibody (HY-P80976) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of HepG2 cells labeling BRD4 with BRD4 antibody (HY-P80976) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with BRD4 antibody (HY-P80976) at 1/200 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Rat brain tissue using BRD4 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80976, 1/100) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Rat brain tissue using BRD4 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80976, 1/100) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-P, ICC/IF, IP

Liquid

Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40%Glycerol, 0.01% sodium azide and 0.05% BSA.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

A synthesized peptide derived from human Brd4