重组Anti-TPX2抗体[EPR23180-4] (ab252945)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23180-4] to TPX2
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IP
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-TPX2抗体[EPR23180-4]
参阅全部 TPX2 一抗 -
描述
兔单克隆抗体[EPR23180-4] to TPX2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details -
种属反应性
与反应: 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: Raji, HEK-293 and HeLa whole cell lysate. IHC-P: Human lung cancer, breast cancer and colon cancer tissue. ICC/IF: HeLa and HEK-293 cells. Flow Cyt (intra): HeLa and Raji cells. IP: Raji whole cell lysate.
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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 +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: 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
EPR23180-4 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab252945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) |
1/500.
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ICC/IF |
1/100.
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WB |
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 86 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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说明 |
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Flow Cyt (Intra)
1/500. |
ICC/IF
1/100. |
WB
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 86 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
靶标
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功能
Spindle assembly factor. Required for normal assembly of mitotic spindles. Required for normal assembly of microtubules during apoptosis. Required for chromatin and/or kinetochore dependent microtubule nucleation. Mediates AURKA localization to spindle microtubules. Activates AURKA by promoting its autophosphorylation at 'Thr-288' and protects this residue against dephosphorylation. -
组织特异性
Expressed in lung carcinoma cell lines but not in normal lung tissues. -
序列相似性
Belongs to the TPX2 family. -
发展阶段
Exclusively expressed in proliferating cells from the transition G1/S until the end of cytokinesis. -
翻译后修饰
Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Nucleus. Cytoplasm > cytoskeleton > spindle. Cytoplasm > cytoskeleton > spindle pole. During mitosis it is strictly associated with the spindle pole and with the mitotic spindle, whereas during S and G2, it is diffusely distributed throughout the nucleus. Is released from the nucleus in apoptotic cells and is detected on apoptotic microtubules. - Information by UniProt
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数据库链接
- Entrez Gene: 22974 Human
- Omim: 605917 Human
- SwissProt: Q9ULW0 Human
- Unigene: 244580 Human
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别名
- C20ORF1 antibody
- C20orf2 antibody
- Chromosome 20 Open Reading Frame 1 antibody
see all
图片
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All lanes : Anti-TPX2 antibody [EPR23180-4] (ab252945) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 3 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 86 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:25239289, 16489064).
Lysates should be made freshly and used in WB immediately to minimize protein degradation.
Exposure time: 5 secs.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling TPX2 with ab252945 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling TPX2 with ab252945 at 1/4000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of human colon cancer is observed(PMID:23634259, 22207630). The section was incubated with ab252945 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 Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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TPX2 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab252945 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Raji whole cell lysate 10 µg.
Lane 2: ab252945 IP in Raji whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252945 in Raji whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 41 secs.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling TPX2 with ab252945 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TPX2 with ab252945 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 cells labeling TPX2 with ab252945 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in HEK-293 cells. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling TPX2 with ab252945 at 1/4000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of human breast cancer is observed (PMID:31494045). The section was incubated with ab252945 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 Goat Anti-Rabbit IgG H&L (HRP).
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 lung cancer tissue labeling TPX2 with ab252945 at 1/4000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of human lung cancer tissue is observed (PMID:23634259, 16489064). The section was incubated with ab252945 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 Goat Anti-Rabbit IgG H&L (HRP).
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)
ab252945 被引用在 1 文献中.
- Wang M et al. miR-548a-3p Weakens the Tumorigenesis of Colon Cancer Through Targeting TPX2. Cancer Biother Radiopharm N/A:N/A (2020). PubMed: 32857606