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  1. Products /
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  3. PathScan® RP BRD4 Isoform C Sandwich ELISA Kit

PathScan® RP BRD4 Isoform C Sandwich ELISA Kit #86520

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  • ELISA

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

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    Supporting Data

    REACTIVITY H
    Application Key:
    • ELISA-ELISA 
    Species Cross-Reactivity Key:
    • H-Human 

    Product Information

    Product Description

    The rapid protocol (RP) PathScan® RP BRD4 Isoform C Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of BRD4 isoform C protein in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with BRD4 isoform C in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of BRD4 isoform C protein. Learn more about your ELISA kit options here.

    *Antibodies in this kit are custom formulations specific to kit.

    Protocol

    Specificity / Sensitivity

    The PathScan® RP BRD4 Isoform C Sandwich ELISA Kit detects endogenous levels of BRD4 isoform C protein. The kit sensitivity is shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

    Species Reactivity:

    Human

    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


    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    For Research Use Only. Not For Use In Diagnostic Procedures.
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