Recombinant Anti-TPX2 antibody [EPR23182-47] (ab252944)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23182-47] to TPX2
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-TPX2 antibody [EPR23182-47]
See all TPX2 primary antibodies -
Description
Rabbit monoclonal [EPR23182-47] to TPX2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293, RAW264.7, L6 and Raji whole cell lysate. IHC-P: Human colon cancer, lung cancer, mouse and rat colon tissue. ICC/IF: RAW264.7 and HeLa cells. Flow Cyt (intra): HeLa cells.
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23182-47 -
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 ab252944 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 |
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Flow Cyt (Intra) |
1/50.
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WB |
1/1000. Predicted molecular weight: 86 kDa.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/50.
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Notes |
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Flow Cyt (Intra)
1/50. |
WB
1/1000. Predicted molecular weight: 86 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/50. |
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
- Entrez Gene: 72119 Mouse
- Entrez Gene: 311546 Rat
- Omim: 605917 Human
- SwissProt: Q9ULW0 Human
- SwissProt: A2APB8 Mouse
- Unigene: 244580 Human
- Unigene: 407737 Mouse
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Alternative names
- C20ORF1 antibody
- C20orf2 antibody
- Chromosome 20 Open Reading Frame 1 antibody
see all
Images
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling TPX2 with ab252944 at 1/50 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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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labeling TPX2 with ab252944 at 1/50 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 RAW 264.7 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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All lanes : Anti-TPX2 antibody [EPR23182-47] (ab252944) at 1/1000 dilution
Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 2 : L6 (rat skeletal muscle myoblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 86 kDaBlocking 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).
The lower bands are mainly caused by degradation (PMID:16287863).
Lysates should be made freshly and used in WB immediately to minimize protein degradation.
Exposure time: 26 secs.
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Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling TPX2 with ab252944 at 1/1000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of mouse colon (PMID:23634259). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).
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Intracellular flow cytometric analysis of 80% methanol fixed 0.1% Tween-20 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TPX2 with ab252944 at 1/50 (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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All lanes : Anti-TPX2 antibody [EPR23182-47] (ab252944) 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 1/50000 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: 3 secs.
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Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling TPX2 with ab252944 at 1/1000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of rat colon (PMID:23634259). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TPX2 with ab252944 at 1/1000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of human lung cancer (PMID:23634259, 16489064). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling TPX2 with ab252944 at 1/1000 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in proliferative cells of human colon cancer (PMID:23634259, 22207630). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).
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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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab252944 has been referenced in 1 publication.
- Cui S & Li C RHPN1-AS1 promotes ovarian carcinogenesis by sponging miR-485-5p and releasing TPX2 mRNA. Oncol Rep 45:N/A (2021). PubMed: 33907841