BromoTag Antibody - BSA Free

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.

• Reactivity: All-NA
• Applications: WB
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

BromoTag Antibody - BSA Free Summary

• Description: NBP3-17999 can be used to detect BromoTag(R) AGB1 (Tocris Catalog # 7686)-induced degradation of Brd4BD2 L387A fusion proteins.
• 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
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Notes

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

Alternate Names for BromoTag Antibody - BSA Free

• Brd4 bromodomain L387A variant
• Brd4BD2L387A
• BromoTag degron

Background

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

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.

Publications for BromoTag Antibody (NBP3-17999)(2)

Publications using NBP3-17999

• Hsia O, Hinterndorfer M, Cowan A et al. An intramolecular bivalent degrader glues an intrinsic BRD4-DCAF16 interaction bioRxiv 2023-02-15 (WB, Human)
• Hsia, O;Hinterndorfer, M;Cowan, AD;Iso, K;Ishida, T;Sundaramoorthy, R;Nakasone, MA;Imrichova, H;Schätz, C;Rukavina, A;Husnjak, K;Wegner, M;Correa-Sáez, A;Craigon, C;Casement, R;Maniaci, C;Testa, A;Kaulich, M;Dikic, I;Winter, GE;Ciulli, A; Targeted protein degradation via intramolecular bivalent glues Nature 2024-02-21 [PMID: 38383787]