Invitrogen

BRD4 Polyclonal Antibody

Product Image Gallery

BRD4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of HeLa cells using an anti-BRD4 polyclonal antibody (Product # PA5-41550). Primary Antibody Dilution: 4 µg/mL; Secondary Antibody: Anti-rabbit Alexa 546; Secondary Antibody Dilution: 2 µg/mL.

BRD4 Antibody in Western Blot (WB) — Western blot analysis of BRD4. 50 µg total protein lysate from human embryonic kidney HEK293T cells (lane 2) and mouse myoblast C2C12 cells (lane 3) in 1X NuPAGE® LDS Sample Buffer (Product # NP0007) + 1mM DTT loaded onto a NuPAGE™ Novex™ 4-12% Bis-Tris protein gel (Product # NP0321PK2). Molecular weight marker loaded for size reference (lane 1). Gel run in 1X NuPAGE® MOPS SDS Running Buffer (Product # NP0001) at room temperature and then transferred to a methanol-activated Novex™ PVDF membrane (Product # LC2002) using a Invitrogen Mini Blot Module (Product # B1000) in 1X NuPAGE® Transfer Buffer (Product # NP0006) at 20V for 2 hours at room temperature. Membrane was rinsed in 1X TBS-T and incubated in blocking solution (5% milk in 1X TBS-T) for 1 hour at room temperature on a rocking platform. HDAC1 detection was performed by incubating blot with rabbit polyclonal anti-BRD4 antibody (Product # PA5-41550; 0.9 mg/mL) diluted at 1:1000 in blocking solution for 1 hour at room temperature on a rocking platform. Primary antibody was washed 10 times with 1X TBS-T over the course of 1 hour at room temperature on a rocking platform. Secondary antibody binding was performed by incubation with goat anti-rabbit IgG-HRP secondary diluted at 1:5000 in blocking solution for 1 hour at room temperature on a rocking platform. Following washes (as above), chemiluminescent imaging was performed. Data courtesy of Dr. Raj Jain's laboratory.

BRD4 Antibody in ChIP assay (CHIP) — ChIP assay analysis of HCT116 cells using an anti-BRD4 polyclonal antibody (Product # PA5-41550).

Product Details

PA5-41550

Applications	Tested Dilution	Publications
Western Blot (WB)	0.2-1 µg/mL	-
Immunocytochemistry (ICC/IF)	4 µg/mL	-
ChIP assay (ChIP)	0.5 µg	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human
Published species	Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic peptide directed towards the C-terminal of human BRD4 View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Affinity Chromatography
Storage buffer	PBS with 2% sucrose
Contains	0.09% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2610136

Product Specific Information

Peptide sequence: EIEIDFETLK PSTLRELERY VTSCLRKKRK PQAEKVDVIA GSSKMKGFSS

Sequence homology: Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%

Target Information

BRD4, a ubiquitously expressed nuclear phosphoprotein, is a member of BET protein family. It is a human homolog of mouse MCAP and consists of 2 N-terminal bromodomains, a central ET-domain, and several kinase-like motifs in its N-terminus. It is expressed as 2 isoforms; the longer variant has a C-terminus region with a praline and glutamine-rich domains. BRD4 is a chromatin-binding protein whose expression is induced in response to growth stimuli. BRD4 acts at different stages of the cell cycle by interacting with proteins including RFC and SPA-1 and regulates cell growth. It also has a role in regulating chromosomal dynamics during mitosis. BRD4 functions as a cellular target for E2 protein of papillomavirus.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

The bromodomain inhibitor JQ1+ reduces calcium-sensing receptor activity in pituitary cell lines.

Journal of molecular endocrinology

Lines KE,Gluck AK,Thongjuea S,Bountra C,Thakker RV,Gorvin CM

PA5-41550 was used in ChIP assay to demonstrate if reducing calcium-sensing receptor expression may be a viable option in the treatment of pituitary tumours.

Mon Jul 26 00:00:00 EDT 2021

Bromodomain and Extraterminal Inhibition by JQ1 Produces Divergent Transcriptional Regulation of Suppressors of Cytokine Signaling Genes in Adipocytes.

Endocrinology

Mota de Sá P,Richard AJ,Stephens JM

PA5-41550 was used in ChIP assay to demonstrate substantially different transcriptional regulation of Socs3 and Cish and suggest distinct roles in adipocytes for these 2 closely related proteins.