BRD2 Antibody (JG40-42)

Western Blot: BRD2 Antibody (JG40-42) [NBP2-75422] - Analysis of BRD2 on SiHa cell lysate using anti-BRD2 antibody at 1/2,000 dilution.

Immunocytochemistry/Immunofluorescence: BRD2 Antibody (JG40-42) [NBP2-75422] - Staining BRD2 in SiHa cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunohistochemistry-Paraffin: BRD2 Antibody (JG40-42) [NBP2-75422] - Analysis of paraffin-embedded human gallbladder tissue using anti-BRD2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow Cytometry: BRD2 Antibody (JG40-42) [NBP2-75422] - Analysis of 293T cells with BRD2 antibody at 1/100 dilution (purple) compared with an unlabelled control (cells without incubation with primary antibody; yellow). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.

Immunocytochemistry/Immunofluorescence: BRD2 Antibody (JG40-42) [NBP2-75422] - Staining BRD2 in 293T cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunocytochemistry/Immunofluorescence: BRD2 Antibody (JG40-42) [NBP2-75422] - Staining BRD2 in A431 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunohistochemistry-Paraffin: BRD2 Antibody (JG40-42) [NBP2-75422] - Analysis of paraffin-embedded human tonsil tissue using anti-BRD2 antibody. Counter stained with hematoxylin.

Immunohistochemistry-Paraffin: BRD2 Antibody (JG40-42) [NBP2-75422] - Analysis of paraffin-embedded human colon tissue using anti-BRD2 antibody. Counter stained with hematoxylin.

Immunohistochemistry-Paraffin: BRD2 Antibody (JG40-42) [NBP2-75422] - Analysis of paraffin-embedded rat colon tissue using anti-BRD2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Product Discontinued

BRD2 Antibody (JG40-42) Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Synthetic peptide within Human BRD2 aa 1-50 / 801. (SwissProt: P25440 Human)
• Localization: Nucleus.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: BRD2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:100
  • Immunocytochemistry/ Immunofluorescence 1:50-1:200
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:100-1:400
  • Western Blot 1:500-1:2000
• Theoretical MW: 88 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: TBS (pH7.4), 0.05% BSA, 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for BRD2 Antibody (JG40-42)

• BRD2
• bromodomain containing 2
• bromodomain-containing 2
• bromodomain-containing protein 2
• D6S113E
• female sterile homeotic-related gene 1
• FSRG1
• FSRG1FSH
• KIAA9001FLJ31942
• NAT
• O27.1.1
• Really interesting new gene 3 protein
• RING3
• RING3DKFZp686N0336
• RNF3

Background

BRD2 (bromodomain-containing 2) is a nuclear mitogen-activated kinase that plays a role in transcription of cell-cycle-regulated genes (1). Alternate names for BRD2 include RING3, NAT, RNF3, FSRG1, D6S113E, FLJ31942, DKFZp686N0336, and KIAA9001 (2). BRD2 belongs to the BET (bromodomain/extra terminal domain) family of proteins which contain two tandem bromodomains (BDI and BDII) and an extra-terminal (ET) domain (1, 3). In mammals there are four BET paralogs (BRD2, BRD3, BRD4, and BRDT) which have a similar amino acid sequence, domain organization, and function (3). The bromodomain of BRD2 is a ~110 amino acids conserved sequence that forms 4 alpha-helices (alphaZ, alphaA, alphaB, and alphaC) and 2 loops (ZA and BC) that can bind to the acetylated lysine-12 residue of chromatin histone H4 and is required for epigenetic regulation of gene transcription (1-3). The ET is a conserved region of ~80 amino acids that recruits specific effector proteins (3). The theoretical molecular weight of BRD2 is 88 kDa but the observed band is typically ~110 kDa due to post-translational modifications. The coding region of the Brd2 gene contains 11 exons and covers over 6 kb of DNA (3). Furthermore, the gene maps to the major histocompatibility complex (MHC) class II region on chromosome 6p21.3 (2).

BRD2 and the other BET proteins have been implicated in a variety of diseases and pathologies. The BET proteins are known drivers of cancer through mutation and over-expression (1). Recently, in studies examining the role of Type 2 diabetes and obesity in breast cancer progression, the BET proteins have been shown to be critical regulators of metabolism and metastasis and are co-activators for the transcription of genes that encode pro-inflammatory cytokines in immune cells infiltrating the breast cancer microenvironment (1). Accordingly, knockdown of Brd2 in mice protected the animals from developing Type 2 diabetes and stopped the inflammatory response typically elicited by obesity (4). BRD2 is also highly expressed in the brain and the gene has been shown to play a role in juvenile myoclonic epilepsy, a common form of epilepsy that typically reveals itself during adolescence (5). In addition to the brain, BRD2 is highly expressed in the bone marrow and consequently its kinase activity has been shown to increase upon cellular proliferation and is significantly elevated in the peripheral blood lymphocytes of lymphoma patients (2, 3).

Research has been done to better understand protein interactions with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of the novel coronavirus disease 2019 (COVID-19), as possible targets for drug therapies. It was recently described that that the transmembrane envelope protein (E) of SARS-CoV-2 binds to both BRD2 and BRD4, suggesting that bromodomain inhibitors could be a potential drug target (6). More specifically, the bromodomain inhibitors could be relevant regarding the secondary immune-related consequences that arise from SARS-CoV-2 infection (6). Bromodomain inhibitors are currently the focus of multiple clinical trials as a potential therapeutic in cancer and pulmonary arterial hypertension (6).

References

1. Andrieu, G.P., Shafran, J.S., Deeney, J.T., Bharadwaj, K.R., Rangarajan, A., & Denis, G.V. (2018). BET proteins in abnormal metabolism, inflammation, and the breast cancer microenvironment. J Leukoc Biol. https://doi:10.1002/JLB.5RI0917-380RR

2. BRD2 bromodomain 2 (human), NCBI

3. Taniguchi, Y. (2016). The Bromodomain and Extra-Terminal Domain (BET) Family: Functional Anatomy of BET Paralogous Proteins. Int J Mol Sci. https://doi:10.3390/ijms17111849

4. Wang, F., Deeney, J.T., & Denis, G.V. (2013). Brd2 gene disruption causes "metabolically healthy" obesity: epigenetic and chromatin-based mechanisms that uncouple obesity from type 2 diabetes. Vitam Horm. https://doi:10.1016/B978-0-12-407766-9.00003-1

5. Gilsoul, M., Grisar, T., Delgado-Escueta, A.V., de Nijs, L., & Lakaye, B. (2019). Subtle Brain Developmental Abnormalities in the Pathogenesis of Juvenile Myoclonic Epilepsy. Front Cell Neurosci. https://doi:10.3389/fncel.2019.00433

6. Harrison, C. (2020). Drug researchers pursue new lines of attack against COVID-19. Nat Biotechnol. https://doi.org/10.1038/d41587-020-00013-z

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.

Bioinformatics

• Gene Symbol: BRD2