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Key Product Details

Species Reactivity

Validated:

Non-species specific

Cited:

Human

Applications

Validated:

Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep

Concentration

1.0 mg/ml

Product Specifications

Immunogen

Partial recombinant protein made to the second bromodomain of Brd4 with a L387A mutation (see Bond, et al PMID 34652918).

Clonality

Polyclonal

Host

Sheep

Description

NBP3-17999 can be used to detect BromoTag(R) AGB1 (Tocris Catalog # 7686)-induced degradation of Brd4BD2 L387A fusion proteins.

Scientific Data Images for BromoTag Antibody

Western Blot: BromoTag Antibody [NBP3-17999]

Western Blot: BromoTag Antibody [NBP3-17999]

Western Blot: BromoTag Antibody [NBP3-17999] - Testing of NBP3-17999 antibody on a BromoTag containing endogenous heterozygous HEK293 cell line, against a well-characterized commercial Brd2 antibody. BromoTag contains Brd4L387A mutation that was used as the original immunogen. Total protein ranged from 2.5-40 ug per well. A HEK293 WT cell line was used as a control. Ponceaus Stain was used as a loading control. NBP3-17999 is demonstrated to be a selective BromoTag antibody.

Applications for BromoTag Antibody

Application
Recommended Usage

Western Blot

Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: BromoTag

BromoTag is a novel TAG degradation platform designed to degrade intracellular proteins of interest. The BromoTag system requires the target protein to be expressed as a fusion protein with a TAG protein, called a degron tag. The BromoTag consists of a bromo and extracellular terminal (BET) bromodomain Brd4 variant L387A as the degron tag. The tag mediates formation of a tertiary complex between the von Hippel Lindau (VHL) E3 ligase and the fusion protein. The fusion protein becomes ubiquitinated by the E3 ligase, leading to proteasomal degradation of the target protein and degrader BromoTag recycling. An advantage of this system is that the degron tag removes the requirement for proteolysis targeting chimeras (PROTACs) or binding ligands for the protein of interest to recruit the E3 ubiquitin ligase. Degradation by BromoTag is an alternative strategy compared to conventional genetic knockout or knockdown. Additionally, the dosage can be modified to control the level of protein degradation and knockdown. The Bromotag system is also reversible, where in cell culture conditions the degrader can be washed out from the media.

BromoTag(R) is a registered trademark of the University of Dundee and is used under license.

References

Bond AG, Craigon C, Chan KH, et al. Development of BromoTag: A "Bump-and-Hole"-PROTAC System to Induce Potent, Rapid, and Selective Degradation of Tagged Target Proteins. J Med Chem. 2021;64(20):15477-15502. https://doi.org/10.1021/acs.jmedchem.1c01532

Alternate Names

Brd4 bromodomain L387A variant, Brd4BD2L387A, BromoTag degron

Additional BromoTag Products

Product Documents for BromoTag Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for BromoTag Antibody

BromoTag(R) is a registered trademark of the University of Dundee and is used under license.

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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