Name: Anti-BRD3 antibody [2088C3a]
SKU: ab50818
Rating: 4 (1 reviews)

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Key features and details

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

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Primary

Protein

Secondary

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Primary - Mouse Anti-BRD3 antibody [2088C3a] (ab50818)
Flow Cyt, WB, IP, ICC/IF

Protein - Recombinant Human BRD3 protein (ab187453)
SDS-PAGE, MS

Secondary - Goat Anti-Mouse IgG H&L (HRP) (ab205719)
WB, IP, ELISA, IHC-P

View more associated products

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: 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 9^th 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.

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...
Purity

Protein G purified
Purification notes

Purified from TCS
Clonality

Monoclonal
Clone number

2088C3a
Isotype

IgG1
Research areas

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
KO cell lines
- Human BRD3 knockout HEK293T cell line (ab266793)
KO cell lysates
- Human BRD3 knockout HEK293T cell lysate (ab258335)
Positive Controls
- HeLa whole cell lysate (ab29545)
- NIH 3T3 whole cell lysate (ab7179)
Recombinant Protein
- Recombinant Human BRD3 protein (ab187453)

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		1/50. Detects a band of approximately 90 kDa (predicted molecular weight: 80 kDa).
IP		Use at 5-1 µg/mg of lysate.
ICC/IF		1/10.

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.
Target information above from: UniProt accession Q15059 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 8019 Human
- Omim: 601541 Human
- SwissProt: Q15059 Human
- Unigene: 522472 Human
- Unigene: 654869 Human
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

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 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)

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)

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)

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)

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 Cytometric analysis using ab50818 (0.5ug). Secondary antibody used: AlexaFluor®488 Goat Anti-mouse IgG (0.25 µg)

Flow cytometry protocols
Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet

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

ab50818 has been referenced in 7 publications.

Lambert JP et al. Interactome Rewiring Following Pharmacological Targeting of BET Bromodomains. Mol Cell 73:621-638.e17 (2019). PubMed: 30554943
Wessel SR et al. Functional Analysis of the Replication Fork Proteome Identifies BET Proteins as PCNA Regulators. Cell Rep 28:3497-3509.e4 (2019). PubMed: 31553917
Ebine K et al. Interplay between interferon regulatory factor 1 and BRD4 in the regulation of PD-L1 in pancreatic stellate cells. Sci Rep 8:13225 (2018). PubMed: 30185888
Chan KH et al. Impact of Target Warhead and Linkage Vector on Inducing Protein Degradation: Comparison of Bromodomain and Extra-Terminal (BET) Degraders Derived from Triazolodiazepine (JQ1) and Tetrahydroquinoline (I-BET726) BET Inhibitor Scaffolds. J Med Chem 61:504-513 (2018). PubMed: 28595007
Gadd MS et al. Structural basis of PROTAC cooperative recognition for selective protein degradation. Nat Chem Biol 13:514-521 (2017). PubMed: 28288108
Kumar K et al. BET inhibitors block pancreatic stellate cell collagen I production and attenuate fibrosis in vivo. JCI Insight 2:e88032 (2017). PubMed: 28194432
Tögel L et al. Dual Targeting of Bromodomain and Extraterminal Domain Proteins, and WNT or MAPK Signaling, Inhibits c-MYC Expression and Proliferation of Colorectal Cancer Cells. Mol Cancer Ther 15:1217-26 (2016). PubMed: 26983878

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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

Abcam user community

Verified customer

Submitted Jan 29 2013

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Purification notes

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Applications

Target

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (7)

Customer reviews and Q&As

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