重组Anti-BRD2抗体[EPR7642] - ChIP Grade (ab139690)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7642] to BRD2 - ChIP Grade
- Suitable for: WB, ICC/IF, IHC-P, Flow Cyt (Intra), ChIP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-BRD2抗体[EPR7642] - ChIP Grade
参阅全部 BRD2 一抗 -
描述
兔单克隆抗体[EPR7642] to BRD2 - ChIP Grade -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IHC-P, Flow Cyt (Intra), ChIPmore details
不适用于: IP -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human BRD2 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P25440 -
阳性对照
- WB: HEK293T, Jurkat, MOLT4, NCCIT and HeLa whole cell lysate (ab150035). ICC/IF: HeLa and wild-type HAP1 cells. IHC-P: Human testis tissue. ChIP: Nuclear extract from LNCaP cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at -20ºC. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR7642 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-BRD2 antibody [EPR7642] (ab197865)
- Alexa Fluor® 647 Anti-BRD2 antibody [EPR7642] (ab197866)
- HRP Anti-BRD2 antibody [EPR7642] (ab198536)
- Alexa Fluor® 555 Anti-BRD2 antibody [EPR7642] (ab213347)
- Alexa Fluor® 594 Anti-BRD2 antibody [EPR7642] (ab215507)
- Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab139690于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (2) |
1/1000 - 1/10000. Predicted molecular weight: 88 kDa.
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ICC/IF |
Use a concentration of 0.5 µg/ml.
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IHC-P | (2) |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ChIP |
Use at an assay dependent concentration.
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说明 |
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WB
1/1000 - 1/10000. Predicted molecular weight: 88 kDa. |
ICC/IF
Use a concentration of 0.5 µg/ml. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIP
Use at an assay dependent concentration. |
靶标
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功能
May play a role in spermatogenesis or folliculogenesis. -
序列相似性
Contains 2 bromo domains.
Contains 1 ET domain. -
结构域
One bromodomain is sufficient for a partial interaction with histone H4 acetylated at 'Lys-13'. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 6046 Human
- Omim: 601540 Human
- SwissProt: P25440 Human
- Unigene: 75243 Human
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别名
- BRD 2 antibody
- Brd2 antibody
- BRD2_HUMAN antibody
see all
图片
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All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690) at 1/1000 dilution
Lane 1 : Wild-type A549 ab288558 cytoplasm cell lysate
Lane 2 : Wild-type A549 ab288558 nuclear cell lysate
Lane 3 : BRD2 knockout A549 C7 cytoplasm cell lysate
Lane 4 : BRD2 knockout A549 C7 nuclear cell lysate
Lane 5 : Wild-type HEK-293T ab255553 cell lysate
Lane 6 : BRD2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 88 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-BRD2 antibody [EPR7642] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse Anti-Nucleophosmin antibody [FC82291] (ab10530) loading control staining at 2 ug/mL imaged in ECL. In Western blot, ab139690 was shown to bind specifically to BRD2. A band was observed at 110 kDa in wild-type A549 cell lysates with no signal observed at this size in BRD2 knockout cell line. To generate this image, wild-type and BRD2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween®20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 20 seconds exposure time. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and HRP conjugated Goat anti-Mouse (H+L) at 1/20000 dilution.
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All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : BRD2 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 88 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab139690 observed at 110 kDa. Red - loading control ab8245 observed at 36 kDa.
ab139690 Anti-BRD2 antibody [EPR7642] was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate ab257191) was used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab139690 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Chromatin was prepared from LNCaP cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab139690 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: BRD2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab139690observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.ab139690 was shown to specifically react with BRD2 when BRD2 knockout samples were used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab139690 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690) at 1/1000 dilution
Lane 1 : MOLT4 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : NCCIT cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 88 kDa -
ab139690 staining Brd2 in wild-type HAP1 cells (top panel) and BRD2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab139690 at 0.5μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunofluorescence analysis of HeLa cells labeling BRD2, with ab139690 at 1/250 dilution.
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Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling BRD2 with ab139690 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing HeLa cells stained with ab139690 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab139690, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (24)
ab139690 被引用在 24 文献中.
- Vichaikul S et al. Inhibition of bromodomain extraterminal histone readers alleviates skin fibrosis in experimental models of scleroderma. JCI Insight 7:N/A (2022). PubMed: 35349485
- Deng Y et al. ARV-771 Acts as an Inducer of Cell Cycle Arrest and Apoptosis to Suppress Hepatocellular Carcinoma Progression. Front Pharmacol 13:858901 (2022). PubMed: 35600879
- Chen IP et al. Viral E protein neutralizes BET protein-mediated post-entry antagonism of SARS-CoV-2. Cell Rep 40:111088 (2022). PubMed: 35839775
- Ding D et al. Retinoblastoma protein as an intrinsic BRD4 inhibitor modulates small molecule BET inhibitor sensitivity in cancer. Nat Commun 13:6311 (2022). PubMed: 36274096
- Imaide S et al. Trivalent PROTACs enhance protein degradation via combined avidity and cooperativity. Nat Chem Biol 17:1157-1167 (2021). PubMed: 34675414