重组Anti-CENPB抗体[EPR24047-64] (ab259855)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24047-64] to CENPB
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CENPB抗体[EPR24047-64]
参阅全部 CENPB 一抗 -
描述
兔单克隆抗体[EPR24047-64] to CENPB -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IP, ICC/IF, IHC-P, WBmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Wild-type A431, Jurkat, HeLa, HeLa, HEK-293T, NIH/3T3 and PC-12 cell lysates. IHC-P: Human pancreas, Human gastric cancer and Human ovarian carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt(Intra) : HeLa cells. IP: HeLa and NIH/3T3 cells.
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常规说明
IHC-P, ICC and FC - suitable for human only
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 +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR24047-64 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lysates
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab259855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 |
---|---|---|
Flow Cyt (Intra) |
1/50.
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IP |
1/30.
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ICC/IF |
1/50.
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IHC-P |
1/100.
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WB |
1/1000. Predicted molecular weight: 65 kDa.
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说明 |
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Flow Cyt (Intra)
1/50. |
IP
1/30. |
ICC/IF
1/50. |
IHC-P
1/100. |
WB
1/1000. Predicted molecular weight: 65 kDa. |
靶标
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功能
Interacts with centromeric heterochromatin in chromosomes and binds to a specific subset of alphoid satellite DNA, called the CENP-B box. May organize arrays of centromere satellite DNA into a higher order structure which then directs centromere formation and kinetochore assembly in mammalian chromosomes. -
序列相似性
Contains 1 HTH CENPB-type DNA-binding domain.
Contains 1 HTH psq-type DNA-binding domain. -
细胞定位
Nucleus. Chromosome > centromere. - Information by UniProt
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数据库链接
- Entrez Gene: 1059 Human
- Entrez Gene: 12616 Mouse
- Entrez Gene: 362217 Rat
- Omim: 117140 Human
- SwissProt: P07199 Human
- SwissProt: P27790 Mouse
- Unigene: 516855 Human
- Unigene: 440169 Mouse
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别名
- CENP B antibody
- CENP-B antibody
- CENPB antibody
see all
图片
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling CENPB with ab259855 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 µg/mL dilution (Green). Confocal image showing centromere staining in HeLa cell line. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5µg/mL dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution.
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All lanes : Anti-CENPB antibody [EPR24047-64] (ab259855) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/50000 dilution
Lane 4 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 65 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were made freshly and used in WB test immediately to minimize protein degradation.
Fresh lysates are preferred in this product.
Exposure time: 3 minutes
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All lanes : Anti-CENPB antibody [EPR24047-64] (ab259855) at 1/1000 dilution
Lane 1 : Wild-type A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : CENPB knockout A431 whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Predicted band size: 65 kDaBlocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lanes 1-4: Merged signal (red and green). Green - ab259855 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.
ab259855 Anti-CENPB antibody [EPR24047-64] was shown to specifically react with CENPB in wild-type A431 cells. Loss of signal was observed when the knockout cell line ab274919 (knockout cell lysate ab274977) was used. Wild-type and CENPB knockout samples were subjected to SDS-PAGE. ab259855 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight 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 10000 dilution for 1 hour at room temperature before imaging.
Fresh lysates are preferred in this product.
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CENPB was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 µg with ab259855 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259855 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug
Lane 2: abab259855 IP in NIH/3T3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259855 in NIH/3T3 whole cell lysateBlocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
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CENPB was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab259855 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259855 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: abab259855 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259855 in HeLa whole cell lysateBlocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CENPB with ab259855 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labelling CENPB with ab259855 at 1/100 (5.39 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on centromere in human ovarian carcinoma (PMID: 12839935).The section was incubated with ab259855 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labelling CENPB with ab259855 at 1/100 (5.39 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on centromere in human gastric cancer (PMID: 12839935). The section was incubated with ab259855 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling CENPB with ab259855 at 1/100 (5.39 μg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on centromere in human pancreas (PMID: 12839935). The section was incubated with ab259855 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (1)
ab259855 被引用在 1 文献中.
- Zelceski A et al. MND1 and PSMC3IP control PARP inhibitor sensitivity in mitotic cells. Cell Rep 42:112484 (2023). PubMed: 37163373