Invitrogen

BRD4 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

View all (23) BRD4 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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BRD4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Bromodomain-containing protein 4 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with BRD4 Polyclonal Antibody (Product # PA5-85662) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

BRD4 Antibody in Western Blot (WB) — Western blot was performed using Anti-BRD4 Polyclonal Antibody (Product # PA5-85662) and a 200kDa band corresponding to Bromodomain-containing protein 4 was observed across the cell lines tested. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), K-562 (Lane 2), HEK-293 (Lane 3), U-2 OS (Lane 4), HT-1080 (Lane 5) and C2C12 (Lane 6) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

BRD4 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of the target protein. K-562 cells were transfected with Bromodomain-containing protein 4 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-BRD4 Polyclonal Antibody (Product # PA5-85662). {KD}

Product Details

PA5-85662

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	-
Immunohistochemistry (Paraffin) (IHC (P))	Assay-dependent	-
Immunocytochemistry (ICC/IF)	1:100	-

Product Specifications
Species Reactivity	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein encompassing a sequence within the C-terminus region of human BRD4. View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.46 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol, 1% BSA
Contains	0.025% ProClin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2792801

Product Specific Information

Keep as concentrated solution.

Predicted reactivity: Pig (98%).

Positive Control: 293T nuclear extract.

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

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.

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Bioinformatics

Protein Aliases: BRD4-NUT FUSION; bromodomain-containing 4; bromodomain-containing 5; Bromodomain-containing protein 4; chromosome-associated protein; fsh-like protein; MCAP; Mitotic chromosome-associated protein; Protein HUNK1

Gene Aliases: BRD4; Brd5; CAP; HUNK1; HUNKI; MCAP; WI-11513

UniProt ID: (Human) O60885, (Mouse) Q9ESU6

Entrez Gene ID: (Human) 23476, (Mouse) 57261

Function(s)