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
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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)
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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.
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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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at 4°C (stable for up to 12 months). Store at -20°C or -80°C. -
Storage buffer
pH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 1% BSA, PBS -
Concentration information loading...
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Purity
Protein G purified -
Purification notes
Purified from TCS -
Clonality
Monoclonal -
Clone number
2088C3a -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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.
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ICC/IF |
1/10.
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Notes |
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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
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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
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Database links
- Entrez Gene: 8019 Human
- Entrez Gene: 67382 Mouse
- Entrez Gene: 362092 Rat
- Omim: 601541 Human
- SwissProt: Q15059 Human
- SwissProt: Q8K2F0 Mouse
- Unigene: 522472 Human
- Unigene: 654869 Human
see all -
Alternative names
- BRD 3 antibody
- Brd3 antibody
- BRD3_HUMAN antibody
see all
Images
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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.
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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.
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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? -
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? -
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? -
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 )
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Flow Cytometric analysis using ab50818 (0.5ug). Secondary antibody used: AlexaFluor®488 Goat Anti-mouse IgG (0.25 µg)
Protocols
Datasheets and documents
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Datasheet download
References (12)
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