Human Bromodomain-containing protein 4(BRD4) ELISA kit

Code CSB-EL002802HU
Size 96T,5×96T,10×96T How to order?
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Human BRD4 ELISA Kit was designed for the quantitative measurement of Human BRD4 protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 18.75 pg/mL-1200 pg/mL and the sensitivity is 4.68 pg/mL.

Target Name bromodomain containing 4
Alternative Names Brd4 ELISA Kit; BRD4-NUT FUSION ELISA Kit; BRD4-NUT fusion oncoprotein ELISA Kit; BRD4_HUMAN ELISA Kit; Bromodomain containing 4 ELISA Kit; bromodomain containing protein 4 ELISA Kit; Bromodomain-containing protein 4 ELISA Kit; CAP ELISA Kit; chromosome associated protein ELISA Kit; HUNK1 ELISA Kit; HUNKI ELISA Kit; MCAP ELISA Kit; Mitotic chromosome-associated protein ELISA Kit; Protein HUNK1 ELISA Kit
Abbreviation BRD4
Uniprot No. O60885
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 18.75 pg/mL-1200 pg/mL
Sensitivity 4.68 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human BRD4 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 SampleSerum(n=4)
1:1Average %89
Range %82-95
1:2Average %87
Range %80-93
1:4Average %90
Range %83-98
1:8Average %95
Range %90-102
Recovery
The recovery of human BRD4 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample TypeAverage % RecoveryRange
Serum (n=5) 9792-107
EDTA plasma (n=4)8580-90
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/mlOD1OD2AverageCorrected
12001.963 1.903 1.933 1.779
6001.624 1.573 1.599 1.445
3001.217 1.182 1.200 1.046
1500.952 0.934 0.943 0.789
750.571 0.526 0.549 0.395
37.50.335 0.317 0.326 0.172
18.750.206 0.199 0.203 0.049
00.159 0.148 0.154  
ELISA Data Analysis Watch ELISA data processing video & download Curve Expert if needed
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Customer Reviews and Q&A

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 Q&A
Q:

Customer need to detect the changes in BRD4 protein in mammalian cell lines treated with compound.
1. Can we check the BRD4 epitope that is targeted by the rat mAb and detection Ab, is it close to the N or C terminus?
2. Also, could we get the antibodies separately? If yes, kindly let me know the product number.

A:
Thanks for your inquiry!
CSB-EL002802HU
Sorry we didn't do epitope mapping before, sothe specific recognition sites is not clear.
If you want to test the changes in BRD4 protein in mammalian cell lines treated with compound, in theory it can be detected out if it doesn't affect the antibody's recognition area and conformation. We suggest you do a pretest first.
Sorry we can not sell the antibodies in the kit separately.
Q:

Per customers request, please provide the following information regarding Human Bromodomain-containing protein 4, BRD4 ELISA Kit CSB-EL002802HU
1) What is the Uniprot/Accession number that this kit is designed for? Is the kit designed for XYZ species or is it designed for a different species such as Human and cross reacts with XYZ species?
2) Is the capture antibody a monoclonal or polyclonal?
3) What is the host of the capture antibody?
4) What’s the immunogen or immunogen range of the capture antibody?
5) What’s is the species of the immunogen of the capture antibody, human or something else?
6) Is the detection antibody a monoclonal or polyclonal?
7) What is the host of the detection antibody?
8) What is the immunogen or immunogen range of the detection antibody?
9) What is the species of the immunogen of the detection antibody? Human or something else?
10) What’s is that standard? A recombinant protein or something else? If a recombinant protein, what is the host?
11) If recombinant protein, is it full length or partial?
12) What is the species of the standard– human or something else?
13) What sample type was tested with this kit (serum? plasma? Something else?) and what were the reference values.

A:
Thanks for your inquiry!
CSB-EL002802HU
This kit is designed according to: https://www.uniprot.org/uniprot/O60885. It is designed for human.
Capture antibody is rat monoclonal antibody and immunogen is 293 cell-derived recombinant protein.
Detection antibody is goat polyclonal antibody and immunogen is 293 cell-derived recombinant protein.
Standard is 293 cell-derived recombinant protein.
Sorry we can not disclose the sequence of standard and immunogen.
We tested undiluted serum and plasma samples before and the test value is 20--75pg/ml.
Pls let me know if you have any further questions. Thank you.

Target Background

Function
(From Uniprot)
Chromatin reader protein that recognizes and binds acetylated histones and plays a key role in transmission of epigenetic memory across cell divisions and transcription regulation. Remains associated with acetylated chromatin throughout the entire cell cycle and provides epigenetic memory for postmitotic G1 gene transcription by preserving acetylated chromatin status and maintaining high-order chromatin structure. During interphase, plays a key role in regulating the transcription of signal-inducible genes by associating with the P-TEFb complex and recruiting it to promoters. Also recruits P-TEFb complex to distal enhancers, so called anti-pause enhancers in collaboration with JMJD6. BRD4 and JMJD6 are required to form the transcriptionally active P-TEFb complex by displacing negative regulators such as HEXIM1 and 7SKsnRNA complex from P-TEFb, thereby transforming it into an active form that can then phosphorylate the C-terminal domain (CTD) of RNA polymerase II. Promotes phosphorylation of 'Ser-2' of the C-terminal domain (CTD) of RNA polymerase II. According to a report, directly acts as an atypical protein kinase and mediates phosphorylation of 'Ser-2' of the C-terminal domain (CTD) of RNA polymerase II; these data however need additional evidences in vivo. In addition to acetylated histones, also recognizes and binds acetylated RELA, leading to further recruitment of the P-TEFb complex and subsequent activation of NF-kappa-B. Also acts as a regulator of p53/TP53-mediated transcription: following phosphorylation by CK2, recruited to p53/TP53 specific target promoters.; Acts as a chromatin insulator in the DNA damage response pathway. Inhibits DNA damage response signaling by recruiting the condensin-2 complex to acetylated histones, leading to chromatin structure remodeling, insulating the region from DNA damage response by limiting spreading of histone H2AX/H2A.x phosphorylation.
Gene References into Functions
  1. Genetic and pharmacological inhibition of BRD4 suppressed IL-1beta-induced expression and translocation of HMGB1. Chromatin immunoprecipitation (ChIP) showed the enrichment of BRD4 around the HMGB1 upstream non-promoter region, which diminished with JQ1 treatment. PMID: 28844955
  2. miR204 directly binds to UCA1 and the 3'untranslated region of BRD4. Furthermore, UCA1 competed with BRD4 for miR204 binding. miR204 knockdown enhanced BRD4 expression, which can be partially restored by short hairpinUCA1. PMID: 30015945
  3. High BRD4 expression is associated with preeclampsia. PMID: 29748248
  4. BRD4 silencing was negatively correlated palomid 529-induced apoptosis in the primary human renal cell carcinoma cells and tumor growth in SCID mice. PMID: 30308518
  5. miR-608 inhibits hepatocellular carcinoma cell proliferation possibly via targeting BET family protein BRD4. PMID: 29777702
  6. MS645 blocks BRD4 binding to transcription enhancer/mediator proteins MED1 and YY1 with potency superior to monovalent BET inhibitors, resulting in down-regulation of proinflammatory cytokines and genes for cell-cycle control and DNA damage repair that are largely unaffected by monovalent BrD inhibition. PMID: 30012592
  7. Study shows that BRD4 is significantly highly expressed in gastric cancer patients and cell lines and positively regulates the expression of c-MYC through transcription regulation and epigenetic levels. Functionally, these result demonstrate that the knockdown of BRD4 represses the proliferation and induces the apoptosis of gastric cancer cells through repression of c-MYC. PMID: 28681984
  8. BRD4 hyperphosphorylation is associated with cellular transformation in NUT midline carcinoma PMID: 28630312
  9. Results suggest structure-based drug design of bromodomain-containing protein 4 (Brd4) inhibitors. PMID: 27494802
  10. Cells harboring the fusion gene are selectively sensitive to small-molecule inhibition of protein targets induced by, or bound to, PAX3-FOXO1-occupied super enhancers. Furthermore, PAX3-FOXO1 recruits and requires the BET bromodomain protein BRD4 to function at super enhancers, resulting in a complete dependence on BRD4 and a significant susceptibility to BRD inhibition PMID: 28446439
  11. Our data suggested that downregulation of BRD4 in gallbladder cancer (GBC) cells induced apoptosis by PI3K/AKT pathway. Inhibition of BRD4 expression may be a novel therapeutic strategy for patients with GBC. PMID: 28766687
  12. these results highlighted the significant genetic contribution of the ARID1B variant, rs73013281, to susceptibility for HCC, especially in interaction with physical activity. PMID: 28415703
  13. In pluripotent cells, Brd2-Brd4 occupy Nodal gene regulatory elements (NREs), but only Brd4 is required for pluripotency gene expression. Brd4 downregulation facilitates pluripotent exit and drives enhanced Brd2 NRE occupancy, thereby unveiling a specific function for Brd2 in differentiative Nodal-Smad2 signalling PMID: 28588073
  14. Findings reveal that PCa-associated ERG can interact and co-occupy with BRD4 in the genome, and suggest this druggable interaction is critical for ERG-mediated cell invasion and PCa progression. PMID: 27223260
  15. Data suggest that pharmacological inhibition of BET proteins could be a potential treatment for renal fibrosis. PMID: 27732564
  16. Our data strongly support the use of CCR2 and CD180 mRNAs as whole blood pharmacodynamic (PD)biomarkers for BRD4 inhibitors, especially in situations where paired tumor biopsies are unavailable. In addition, they can be used as tumor-based PD biomarkers for hematologic tumors. PMID: 28073847
  17. Study identified Brd4 as a novel proline hydroxylation substrate with nearly 60% stoichiometry under normoxia states and its hydroxylation level is enzymatically regulated. PMID: 27764789
  18. Further analysis indicated that JQ1 inhibited the recruitment of BDR4 to the promoter complex of the Myc and Ccnd1 genes in rat thyroid follicular PCCL3 cells, resulting in decreased MYC expression at the mRNA and protein levels to inhibit tumor cell proliferation PMID: 27440272
  19. The Brd4 acetyllysine-binding protein of RelA is involved in activation of polyomavirus JC. PMID: 27007123
  20. BRD4 phosphorylation regulates HPV E2-mediated viral transcription and cellular MMP-9 expression PMID: 27477287
  21. conclude that BRD3/4 and the FLT3-TAK1/NF-kB pathways collectively control a set of targets that are critically important for the survival of human MLL-AF9 cells PMID: 29240787
  22. Diverse gastric cancer cell lines of Asian and Brazilian origins differ in BRD4 and c-MYC expression levels and sensitivity to BET inhibitors. PMID: 27259267
  23. Together, dual inhibition of BRD4 and PI3K by SF2523 suppresses human prostate cancer cell growth in vitro and in vivo. PMID: 29133261
  24. peroxisome proliferator activated receptor alpha peroxisome proliferator-activated receptor alpha PPAR-alpha nuclear receptor subfamily 1 group C member 1 peroxisome proliferative activated receptor alpha peroxisome proliferator-activated nuclear receptor alpha variant 3 PMID: 28012209
  25. Brd4 is involved in different steps of the papillomavirus life cycle. (Review) PMID: 27965149
  26. BRD4 and MYC are essential for the expression of a subgroup of genes induced by class-I HDAC inhibitors. PMID: 28369619
  27. Data show that BRD4 controls RUNX2 by binding to the enhancers (ENHs) and each RUNX2 ENH is potentially controlled by a distinct set of TFs and c-JUN as the principal pivot of this regulatory platform. PMID: 28981843
  28. any of the molecules or processes in the network could be targeted to curb the oncogenic effects of c-Myc, just as BRD4 can be targeted. And since the targeting of metabolic enzymes has proved effective in mouse tumor models, it might be possible to develop new therapies based on the fact that c-Myc has a role in controlling cellular metabolism PMID: 28100396
  29. Brd4 inhibition attenuates unilateral ureteral obstruction-induced fibrosis by blocking TGF-beta-mediated Nox4 expression. PMID: 28063381
  30. The response of the kinome to targeted BETi treatment in a panel of BRD4-dependent ovarian carcinoma (OC) cell lines. PMID: 27452461
  31. A miR-9 mimic represses stimulus-dependent targeting of BRD4. PMID: 27425608
  32. BRD4 and CDK9 have independent, coordinated roles in promoting the myofibroblast transition PMID: 28182006
  33. Long-term treatment with bromodomain-containing protein 4 (BRD4) inhibitors caused telomere shortening in both mouse and human cells, suggesting BRD4 plays a role in telomere maintenance in vivo. PMID: 28854735
  34. The short isoform of BRD4 cooperates with SWI/SNF nucleosome remodelers to repress HIV transcription during latency, a phenotype reversed by BET inhibitor treatment. PMID: 28844864
  35. results indicated that in MPNST samples BRD4 mRNA levels were not upregulated and that MPNST cell lines were relatively insensitive to the bromodomain inhibitor JQ1. PMID: 28813519
  36. High BRD4 expression is associated with stomach neoplasms. PMID: 28481868
  37. These findings reveal a novel mechanism by which the HBV genome hijacks the host P-TEFb-containing complexes to promote its own transcription; both the super elongation complex and BRD4 can bind to the HBV genome. PMID: 28694331
  38. Chemical degrader of BET bromodomain proteins, dBET6, led to the unexpected identification of BRD4 as master regulator of global transcription elongation in acute T-cell leukemia. BRD4 loss does not directly affect CDK9 localization. PMID: 28673542
  39. BRD4 binds and stays associated with chromatin during mitosis, bookmarking early G1 genes and reactivating transcription after mitotic silencing. BRD4 acts as a passive scaffold via its recruitment of vital transcription factors and as an active kinase that phosphorylates RNA polymerase II. A model in which BRD4 actively coordinates chromatin structure and transcription is described. Review. PMID: 27450555
  40. we have discovered that BRD4-bound super-enhancers provide a powerful tool for enriching and prioritizing PC and BC genetic risk loci PMID: 28359301
  41. BRD4 represses autophagy and lysosome gene expression. This repression is alleviated during nutrient deprivation through AMPK-SIRT1 signaling, allowing autophagy activation. BRD4 inhibition enhances autophagic flux and lysosomal function and promotes the degradation of protein aggregates. Sakamaki et PMID: 28525743
  42. The studies show that BRD4 inhibition may have therapeutic implications for the treatment of mitochondrial diseases. PMID: 27666594
  43. A BRD4 missense mutation was the likely disease-causing mutation in this family with autosomal dominant syndromic congenital cataracts associated with neuro-skeletal anomalies. PMID: 28076398
  44. these studies highlight the importance of Brd4 in Helicobacter pylori-induced inflammatory gene expression and suggest that Brd4 could be a potential therapeutic target for the treatment of Helicobacter pylori-triggered inflammatory diseases and cancer PMID: 27084101
  45. BRD4 positively regulates EZH2 transcription through upregulation of C-MYC, and is a novel promising target for pharmacologic treatment in transcriptional program intervention against this intractable disease. PMID: 26939702
  46. We postulate that BRD4-NUT (B4N) complexes override the preexisting histone code with new posttranslational modifications patterns that reflect aberrant transcription and that epigenetically modulate the nucleosome environment toward the NUT-midline carcinoma (NMC) state. PMID: 27698495
  47. The authors show that, upon infection of primary human keratinocytes with human papillomavirus 18 quasivirus, Brd4 activates viral transcription and replication. PMID: 27879331
  48. research reveals that BRD4 probably play a critical role in Renal Cell Carcinoma progression, and is a new promising target for pharmacological treatment directed against this intractable disease. PMID: 28391274
  49. These data provide a detailed mechanism for how activated NF-kappaB and BRD4 control epithelial-mesenchymal transition initiation and transcriptional elongation in model airway epithelial cells in vitro and in a murine pulmonary fibrosis model in vivo. PMID: 27793799
  50. BRD4 localization to lineage-specific enhancers is associated with a distinct transcription factor repertoire. PMID: 27651452

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Involvement in disease A chromosomal aberration involving BRD4 is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. Translocation t(15;19)(q14;p13) with NUT which produces a BRD4-NUT fusion protein.
Subcellular Location Nucleus. Chromosome.; [Isoform B]: Chromosome.
Tissue Specificity Ubiquitously expressed.
Database Links

HGNC: 13575

OMIM: 608749

KEGG: hsa:23476

STRING: 9606.ENSP00000263377

UniGene: Hs.187763

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