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Commit: 56767fe525c928647c8401233a175d0d607d385d
XML generation date: 2024-09-30 01:59:17.283
Product last modified at: 2024-09-30T08:02:41.455Z
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PDP - Template Name: Monoclonal Antibody
PDP - Template ID: *******c5e4b77
  1. Products /
  2. Primary Antibodies /
  3. BRD4 Isoform C (E1Q9N) Rabbit mAb
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

BRD4 Isoform C (E1Q9N) Rabbit mAb #20696

Filter:
  • WB
  • IP
  • IF
  • ChIP

    Supporting Data

    REACTIVITY H M R Mk
    SENSITIVITY Endogenous
    MW (kDa) 105
    Source/Isotype Rabbit IgG
    Application Key:
    • WB-Western Blotting 
    • IP-Immunoprecipitation 
    • IF-Immunofluorescence 
    • ChIP-Chromatin Immunoprecipitation 
    Species Cross-Reactivity Key:
    • H-Human 
    • M-Mouse 
    • R-Rat 
    • Mk-Monkey 

    Product Information

    Product Usage Information

    For optimal ChIP results, use 5 μL of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
    Application Dilution
    Western Blotting 1:1000
    Immunoprecipitation 1:50
    Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
    Chromatin IP 1:100

    Storage

    Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

    Protocol

    Specificity / Sensitivity

    BRD4 Isoform C (E1Q9N) Rabbit mAb recognizes endogenous levels of total BRD4 protein. This antibody does not cross-react with other BRD4 isoforms.

    Species Reactivity:

    Human, Mouse, Rat, Monkey

    Source / Purification

    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu715 of human BRD4 isoform C protein.

    Background

    Bromodomain-containing protein 4 (BRD4) is a member of the bromodomains and extra terminal (BET) family of proteins, which also includes BRD2, BRD3, and BRDT (1-3). BET family proteins contain two tandem bromodomains and an extra terminal (ET) domain, and bind acetyl lysine residues (3). BRD4 is a chromatin-binding protein with a preference for Lys14 on histone H3 as well as Lys5 and Lys12 on histone H4 (4). BRD4 chromatin binding occurs throughout the cell cycle, including condensed mitotic chromosomes, when the majority of genes are silenced (5). BRD4 association with chromatin during mitosis is thought to be an important part of the bookmarking mechanism to accelerate reactivation of the silenced genes upon exit from mitosis (2,6). BRD4 has been shown to facilitate transcription by recruiting the positive transcription elongation factor b (pTEFb) complex that phosphorylates Ser2 of the heptapeptide repeat of the carboxy-terminal domain of RNA polymerase II, promoting transcription elongation (3,7,8). In addition, BRD4 has been found to be part of the super elongation complex and the polymerase associated factor complex (PAFc) in MLL-fusion derived leukemia cell lines, demonstrating a role for BRD4 in the regulation of transcription elongation (9). Research studies have shown that BRD4 (and BET family proteins) may be promising therapeutic targets for various Myc-driven cancers, such as Burkitt’s lymphoma and certain acute myeloid leukemias (1,10,11). Investigators have found molecular inhibition of BET proteins to be effective in inducing apoptosis in various MLL-fusion driven leukemic cell lines by competing BRD3 and BRD4 from chromatin, leading to reduced expression of Bcl-2, Myc, and CDK6 (9). BET inhibition has also been shown to have antitumor activities against nuclear protein in testis (NUT) midline carcinoma cell lines and xenografts in mice where BRD4 is found to be a frequent translocation partner of the NUT protein (12). In addition, BRD4 regulates the expression of some inflammatory genes, and inhibition of BRD4 (and BET family proteins) chromatin binding causes reduced expression of a subset of inflammatory genes in macrophages, leading to protection against endotoxic shock and sepsis (13).

    The expression of BRD4 isoform C, also known as the short isoform (BRD4-S), is similar to the long isoform (BRD4-L). The balance of expression between the two isoforms has been shown to contribute to disease states, where BRD4-S has shown to be oncogenic in a few contexts (14-16). BRD4-S is the predominant isoform that binds to modified histones and has a stronger affinity than BRD4-L (17,18). BRD4-S also plays a role in DNA damage, where it recruits the condensin II complex to control repair (19).
    1. Belkina, A.C. and Denis, G.V. (2012) Nat Rev Cancer 12, 465-77.
    2. Voigt, P. and Reinberg, D. (2011) Genome Biol 12, 133.
    3. Wu, S.Y. and Chiang, C.M. (2007) J Biol Chem 282, 13141-5.
    4. Dey, A. et al. (2003) Proc Natl Acad Sci U S A 100, 8758-63.
    5. Dey, A. et al. (2009) Mol Biol Cell 20, 4899-909.
    6. Zhao, R. et al. (2011) Nat Cell Biol 13, 1295-304.
    7. Jang, M.K. et al. (2005) Mol Cell 19, 523-34.
    8. Yang, Z. et al. (2005) Mol Cell 19, 535-45.
    9. Dawson, M.A. et al. (2011) Nature 478, 529-33.
    10. Muller, S. et al. (2011) Expert Rev Mol Med 13, e29.
    11. Mertz, J.A. et al. (2011) Proc Natl Acad Sci U S A 108, 16669-74.
    12. Filippakopoulos, P. et al. (2010) Nature 468, 1067-73.
    13. Nicodeme, E. et al. (2010) Nature 468, 1119-23.
    14. Wu, S.Y. et al. (2020) Mol Cell 78, 1114-1132.e10.
    15. Crawford, N.P. et al. (2008) Proc Natl Acad Sci U S A 105, 6380-5.
    16. Fernandez, P. et al. (2014) Cell Rep 9, 248-260.
    17. Wang, R. et al. (2012) J Biol Chem 287, 10738-52.
    18. Han, X. et al. (2020) Nat Struct Mol Biol 27, 333-341.
    19. Floyd, S.R. et al. (2013) Nature 498, 246-50.

    Database Links

    UniProt ID: O60885-2


    Entrez-Gene Id: 23476


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