Recombinant Anti-TPX2 antibody [EPR23180-4] (ab252945)
![Western blot - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-9-anti-tpx2-antibody-epr231804-western-blot-lysate.jpg "Western blot - Anti-TPX2 antibody [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
Overview
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Product name
Anti-TPX2 antibody [EPR23180-4]
See all TPX2 primary antibodies -
Description
Rabbit monoclonal [EPR23180-4] to TPX2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23180-4 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab252945 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 (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.
|
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| IP |
1/30.
|
| Notes |
|---|
|
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. |
Target
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Function
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. -
Tissue specificity
Expressed in lung carcinoma cell lines but not in normal lung tissues. -
Sequence similarities
Belongs to the TPX2 family. -
Developmental stage
Exclusively expressed in proliferating cells from the transition G1/S until the end of cytokinesis. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
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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Database links
- Entrez Gene: 22974 Human
- Omim: 605917 Human
- SwissProt: Q9ULW0 Human
- Unigene: 244580 Human
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Alternative names
- C20ORF1 antibody
- C20orf2 antibody
- Chromosome 20 Open Reading Frame 1 antibody
see all
Images
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Western blot - Anti-TPX2 antibody [EPR23180-4] (ab252945)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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![Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-4-anti-tpx2-antibody-epr231804-immunocytochemistry-hela.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-1-anti-tpx2-antibody-epr231804-immunohistochemistry-human-colon-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)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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![Immunoprecipitation - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-8-anti-tpx2-antibody-epr231804-immunoprecipitation-raji.jpg)
Immunoprecipitation - Anti-TPX2 antibody [EPR23180-4] (ab252945)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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![Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-7-anti-tpx2-antibody-epr231804-flow-cytometry-raji.jpg)
Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)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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![Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-6-anti-tpx2-antibody-epr231804-flow-cytometry-hela.jpg)
Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)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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![Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-5-anti-tpx2-antibody-epr231804-immunocytochemistry-hek293.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)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.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-3-anti-tpx2-antibody-epr231804-immunohistochemistry-human-breast-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)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.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-2-anti-tpx2-antibody-epr231804-immunohistochemistry-human-lung-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)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.
-
![Anti-TPX2 antibody [EPR23180-4] (ab252945)](https://www.abcam.com/ps/products/252/ab252945/Images/ab252945-10-benefits-of-recombinant-antibodies.png)
Anti-TPX2 antibody [EPR23180-4] (ab252945)
Protocols
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab252945 has been referenced in 1 publication.
- 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
