BRD4 Antibody

Images

 
Western Blot: BRD4 Antibody [NB100-56487] - Analysis of Brd4 in human HeLa lysate in the 1) absence and 2) presence of immunizing peptide and C) mouse NIH 3T3 lysate using Brd4 antibody at 3 ug/ml. Goat anti-rabbit Ig ...read more
Immunocytochemistry/ Immunofluorescence: BRD4 Antibody [NB100-56487] - BRD4 antibody was tested in Hela cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Simple Western: BRD4 Antibody [NB100-56487] - Simple Western lane view shows a specific band for BRD4 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, PmSpecies Glossary
Applications WB, Simple Western, ICC/IF, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Order Details

BRD4 Antibody Summary

Immunogen
A portion of amino acids 150-200 of human BRD4 was used as the immunogen (NP_490597).
Specificity
The amino acid sequence used as immunogen has 100% identity in human, chimp, mouse and rat. BRD4 is expressed in two forms, the long form is approximately 1400 amino acids and the short form is approximately 722 amino acids.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
BRD4
Purity
Protein G purified
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Applications/Dilutions

Dilutions
  • Western Blot 2-5 ug/ml
  • Simple Western 1:100
  • Immunocytochemistry/Immunofluorescence 10 ug/ml
  • Immunoprecipitation
Application Notes
IP data from customer review. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Positive Control
BRD4 Lysate (NBP2-65723)
Reviewed Applications
Read 2 Reviews rated 3.5
using
NB100-56487 in the following applications:

Reactivity Notes

Chimpanzee.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for BRD4 Antibody

  • bromodomain containing 4
  • bromodomain-containing 4
  • CAPchromosome-associated protein
  • HUNK1bromodomain-containing protein 4
  • HUNKI
  • MCAP
  • Protein HUNK1

Background

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; 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 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for BRD4 Antibody (NB100-56487) (0)

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Reviews for BRD4 Antibody (NB100-56487) (2) 3.52

Average Rating: 3.5
(Based on 2 reviews)
We have 2 reviews tested in 2 species: Human, Rat.

Reviews using NB100-56487:
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IP
(1)
WB
(1)
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Human
(1)
Rat
(1)
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Images Ratings Applications Species Date Details
Immunoprecipitation BRD4 NB100-56487
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reviewed by:
Xiaofeng Wang
IP Human 06/10/2014
View

Summary

ApplicationImmunoprecipitation
Sample TestedBT16 Nuclear Extract
SpeciesHuman
Lot08082688B-02
CommentsThis antibody looks to be suitable for IP, but need to note there are several non-specific bands when doing the IB.

Blocking

Blocking Details10% Non-fat milk in PBST for30 min

Primary Anitbody

Dilution Ratio1:1000; Overnight at 4 degree; in 1% Non-fat milk in PBST

Secondary Antibody

Secondary DescriptionGoat anti-Rabbit IgG-HRP
Secondary Manufacturer Cat#Jackson ImmunoResearch Laboratories
Secondary Concentration1:5000

Details

Detection NotesECL; exposure time: 3 min
Immunoprecipitation DetailsDyna-Protein G beads; 30 ul of beads added; antibody incubation O/N at coldroom, then beads were added for another 3h incubation
Elution DescriptionSDS-Loading buffer @100degree for 5 min

Comments

CommentsThis antibody looks to be suitable for IP, but need to note there are several non-specific bands when doing the IB.
Western Blot BRD4 NB100-56487
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reviewed by:
Matt Stratton
WB Rat 04/19/2014
View

Summary

ApplicationWestern Blot
Sample Tested primary neonatal ventricular cardiomyocytes
SpeciesRat
CommentsTransfer buffer contained 0.1% SDS to assist in transferring high molecular weight proteins form gel to membrane.

Blocking

Blocking Details5% nonfat milk/TBST

Primary Anitbody

Dilution Ratio1:2000 overnight at 4C in 2.5% BSA/TBST

Secondary Antibody

Secondary DescriptionGoat anti-rabbit HRP
Secondary Manufacturer Cat#Southern Biotech 4030-05
Secondary Concentration1:2000

Details

Detection NotesPierce Supersignal West Pico Chemiluminescent Substrate : 3 minute exposure

Comments

CommentsTransfer buffer contained 0.1% SDS to assist in transferring high molecular weight proteins form gel to membrane.

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for BRD4 Antibody (NB100-56487). (Showing 1 - 1 of 1 FAQ).

  1. I would like to know if the antibody NB100-56487 (Brd4) can recognize both the long and short form of Brd4? Based on the datasheet, it seems like this antibody can recognize both forms but I am looking for papers or images that show the ability of the antibody to detect both forms.
    • Based on the sequence used to generate this antibody (NB100-56487), it should detect both the long and short form of the protein. The immunogen falls between amino acids 150-200. Looking at the images available on our website, it looks as though the antibody has a higher affinity for the short protein compared to the long protein. We unfortunately do not have any published references at this time; that utilize our product.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Additional Array Products

Bioinformatics Tool for BRD4 Antibody (NB100-56487)

Discover related pathways, diseases and genes to BRD4 Antibody (NB100-56487). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
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Diseases for BRD4 Antibody (NB100-56487)

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Pathways for BRD4 Antibody (NB100-56487)

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PTMs for BRD4 Antibody (NB100-56487)

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Blogs on BRD4.

Understanding the relationship between NUT and BET proteins in NMC
NUT has been found to fuse with bromodomain-containing proteins 3 and 4 (BRD3 and BRD4) in NUT midline carcinoma (NMC), a very rare, extremely aggressive, and genetically defined human cancer. NMC has recently been designated as a sub classificati...  Read full blog post.

NUT - A Protein Coding Gene
The NUT gene is found on chromosome 15q14 and encodes for the NUT protein which is a key component of the RNA polymerase II Mediator complex. This multi-subunit assembly is required for all RNA pol II-dependent transcriptional activation, coordinating...  Read full blog post.

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Recent Reviews

3.5
5
0
4
1
3
1
2
0
1
0

Xiaofeng Wang
06/10/2014
Application: IP
Species: Human

Matt Stratton
04/19/2014
Application: WB
Species: Rat

Bioinformatics

Gene Symbol BRD4
Entrez
Uniprot