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  1. Link

    tpx2-antibody-epr23180-4-ab252945.pdf

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Cell Biology Cell Cycle Cell Division Spindle
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RecombinantRabMAb

Recombinant Anti-TPX2 antibody [EPR23180-4] (ab252945)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Immunoprecipitation - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • 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.

  • 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.
  • 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
    For more information see here.

    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

  • 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

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Spindle
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Chromosome Structure
    • Centromere
    • Cancer
    • Cell cycle
    • Cell division

Associated products

  • Alternative Versions

    • Anti-TPX2 antibody [EPR23180-4] - BSA and Azide free (ab270612)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

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.
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

  • 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
    modifications

    Phosphorylated 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.
  • Target information above from: UniProt accession Q9ULW0 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 22974 Human
    • Omim: 605917 Human
    • SwissProt: Q9ULW0 Human
    • Unigene: 244580 Human
    • Alternative names

      • C20ORF1 antibody
      • C20orf2 antibody
      • Chromosome 20 Open Reading Frame 1 antibody
      • Differentially expressed in cancerous and non-cancerous lung cells 2 antibody
      • Differentially expressed in cancerous and noncancerous lung cells 2 antibody
      • Differentially expressed in lung cells 2 antibody
      • Differentially expressed in lung cells antibody
      • DIL 2 antibody
      • DIL-2 antibody
      • DIL2 antibody
      • FLS353 antibody
      • GD:C20orf1 antibody
      • HCA 519 antibody
      • HCA519 antibody
      • HCTP 4 antibody
      • HCTP4 antibody
      • Hepatocellular carcinoma associated antigen 519 antibody
      • Hepatocellular carcinoma-associated antigen 519 antibody
      • p100 antibody
      • Preferentially expressed in colorectal cancer antibody
      • Protein fls353 antibody
      • REPP 86 antibody
      • repp86 antibody
      • Restricted expression proliferation associated protein 100 antibody
      • Restricted expression proliferation-associated protein 100 antibody
      • Targeting protein for Xklp2 antibody
      • TPX 2 antibody
      • TPX2 antibody
      • TPX2 microtubule associated homolog antibody
      • TPX2 microtubule associated protein homolog antibody
      • TPX2 microtubule nucleation factor antibody
      • TPX2_HUMAN antibody
      see all

    Images

    • Western blot - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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.

    • Immunoprecipitation - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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.

       

    • Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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. 

       

       

    • Flow Cytometry (Intracellular) - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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. 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-TPX2 antibody [EPR23180-4] (ab252945)
      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)
      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)
      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)
      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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab252945? Please let us know so that we can cite the reference in this datasheet.

    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

    Customer reviews and Q&As

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