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Validated using a knockout cell line

Anti-BRD3 antibody [2088C3a] (ab50818)

Reviews (1) Submit a question References (12)
Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
  • Immunoprecipitation - Anti-BRD3 antibody [2088C3a] (ab50818)
  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRD3 antibody [2088C3a] (ab50818)
  • Flow Cytometry - Anti-BRD3 antibody [2088C3a] (ab50818)

Key features and details

  • Mouse monoclonal [2088C3a] to BRD3
  • Suitable for: Flow Cyt, WB, IP, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-BRD3 antibody [2088C3a]
    See all BRD3 primary antibodies

  • Description

    Mouse monoclonal [2088C3a] to BRD3

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, WB, IP, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment of BRD3 (Human)

  • Positive control

    • WB: HEK293T, Jurkat, HeLa, F2408 and NIH3T3 whole cell lysates. ICC/IF and Flow Cyt: HeLa cells. IP: RIPA lysate of HeLa cells.

  • General notes

    This product was changed from ascites to tissue culture supernatant on 9th August 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab50818 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt
Use a concentration of 0.5 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB (1)
1/50. Detects a band of approximately 90 kDa (predicted molecular weight: 80 kDa).
IP
Use at 1-5 µg/mg of lysate.
ICC/IF
1/10.
Notes
Flow Cyt
Use a concentration of 0.5 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB
1/50. Detects a band of approximately 90 kDa (predicted molecular weight: 80 kDa).
IP
Use at 1-5 µg/mg of lysate.
ICC/IF
1/10.

Target

  • Tissue specificity

    Ubiquitous.

  • Involvement in disease

    Note=A chromosomal aberration involving BRD3 is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. Translocation t(15;9)(q14;q34) with NUT which produces a BRD3-NUT fusion protein.

  • Sequence similarities

    Contains 2 bromo domains.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • BRD 3 antibody
    • Brd3 antibody
    • BRD3_HUMAN antibody
    • Bromodomain containing 3 antibody
    • Bromodomain containing protein 3 antibody
    • Bromodomain-containing protein 3 antibody
    • KIAA0043 antibody
    • Open reading frame X antibody
    • ORFX antibody
    • RING3 like gene antibody
    • RING3-like protein antibody
    • RING3L antibody
    see all

Images

  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    All lanes : Anti-BRD3 antibody [2088C3a] (ab50818) at 1/500 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : BRD3 knockout HEK293T cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Predicted band size: 80 kDa
    Observed band size: 100 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab50818 observed at 100 kDa. Red - loading control ab181602 observed at 36 kDa.

     ab50818 Anti-BRD3 antibody [2088C3a] was shown to specifically react with BRD3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266793 (knockout cell lysate ab258335) was used. Wild-type and BRD3 knockout samples were subjected to SDS-PAGE. ab50818 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-BRD3 antibody [2088C3a] (ab50818)
    Immunoprecipitation - Anti-BRD3 antibody [2088C3a] (ab50818)
    Immunoprecipitation of BRD3 from RIPA lysate of HeLa cells (500µg) using 5µg ab50818. Visualized by western blot using 1/50 ab50518 as primary and anti-mouse IgG secondary (1:2500). Predicted band size 80 KDa.
  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Anti-BRD3 antibody [2088C3a] (ab50818) at 1/200 dilution + F2408 whole cell lysate at 50 µg

    Secondary
    Anti Mouse IgG antibody at 1/2500 dilution

    Predicted band size: 80 kDa
    Observed band size: 100 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Anti-BRD3 antibody [2088C3a] (ab50818) at 1/50 dilution + HeLa whole cell lysate at 50 µg

    Secondary
    Anti-Mouse IgG antibody at 1/2500 dilution

    Developed using the ECL technique.

    Predicted band size: 80 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Western blot - Anti-BRD3 antibody [2088C3a] (ab50818)
    Anti-BRD3 antibody [2088C3a] (ab50818) at 1/50 dilution + NIH3T3 whole cell lysate at 50 µg

    Secondary
    Anti-Mouse IgG antibody at 1/2500 dilution

    Developed using the ECL technique.

    Predicted band size: 80 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-BRD3 antibody [2088C3a] (ab50818)
    Immunocytochemistry/ Immunofluorescence - Anti-BRD3 antibody [2088C3a] (ab50818)
    Immunoflourescent staining of Hela cells using ab50818 Anti-BRD3 Antibody (1/10 dilution) as primary and AlexaFluor®488 Goat Anti-mouse IgG( 1/200 dilution )
  • Flow Cytometry - Anti-BRD3 antibody [2088C3a] (ab50818)
    Flow Cytometry - Anti-BRD3 antibody [2088C3a] (ab50818)
    Flow Cytometric analysis using ab50818 (0.5ug). Secondary antibody used: AlexaFluor®488 Goat Anti-mouse IgG (0.25 µg)

References (12)

Publishing research using ab50818? Please let us know so that we can cite the reference in this datasheet.

ab50818 has been referenced in 12 publications.

  • Zhou X  et al. BET inhibitors combined with chemotherapy synergistically inhibit the growth of NSCLC cells. Oncol Rep 45:N/A (2021). PubMed: 33760217
  • Lam FC  et al. BRD4 prevents the accumulation of R-loops and protects against transcription-replication collision events and DNA damage. Nat Commun 11:4083 (2020). PubMed: 32796829
  • Edwards DS  et al. BRD4 Prevents R-Loop Formation and Transcription-Replication Conflicts by Ensuring Efficient Transcription Elongation. Cell Rep 32:108166 (2020). PubMed: 32966794
  • Ohoka N  et al. Development of Small Molecule Chimeras That Recruit AhR E3 Ligase to Target Proteins. ACS Chem Biol 14:2822-2832 (2019). PubMed: 31580635
  • Lambert JP  et al. Interactome Rewiring Following Pharmacological Targeting of BET Bromodomains. Mol Cell 73:621-638.e17 (2019). PubMed: 30554943
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Western blot abreview for Anti-BRD3 antibody [2088C3a]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEFs)
Loading amount
15 µg
Specification
MEFs
Gel Running Conditions
Reduced Denaturing (4-12)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 29 2013

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