製品の概要

  • 製品名Anti-TPX2 antibody [18D5-1]
    TPX2 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [18D5-1] to TPX2
  • 特異性This antibody detects TPX2.
  • アプリケーション適用あり: ICC/IF, IP, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Recombinant TPX2 protein (Human).

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32795 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent dilution. PubMed: 19910498
IP Use at an assay dependent dilution.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 92 kDa (predicted molecular weight: 86 kDa).
IHC-P Use a concentration of 0.5 - 1 µg/ml.

ターゲット情報

  • 機能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.
  • 組織特異性Expressed in lung carcinoma cell lines but not in normal lung tissues.
  • 配列類似性Belongs to the TPX2 family.
  • 発生段階Exclusively expressed in proliferating cells from the transition G1/S until the end of cytokinesis.
  • 翻訳後修飾Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在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
  • 参照データベース
  • 別名
    • 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_HUMAN antibody
    see all

Anti-TPX2 antibody [18D5-1] 画像

  • ab32795 staining TPX2 in HeLa cells and mouse NIH-3T3 cells (fuzzier pattern, different from the high-quality sharp signal seen in Human cells), by immunofluorescence.
    optimal antibody dilution: 4µg/ml
    optimal fixation protocol: PFA/Triton fixation: 10 min room at room temperature, in 3,7 % PFA diluted in PHEM buffer (45  mM Hepes pH 6,9, 45 mM Pipes pH 6,9, 5 mM MgCl2, 10 mM EGTA) containing 0.2% Triton X-100, followed  by 3 washes in PBS    -  Alternative fixation protocol also gives good staining: 6 min in cold Methanol at -20°C,  then 3 washes in PBS.
    IF was performed following a standard protocol: Blocking, 30 min; primary antibody, 1 hr; secondary antibody, 45 min. All incubations were at 37 °C in PBS/ 0.1% Tween containing 3% BSA.

  • ab32795 (1µg/ml) staining TPX2 in human testis using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and cytoplasmic staining .
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunofluorescent analysis of PLK1 using PLK1 Monoclonal antibody (13E8) ab32795 shows staining in WiDr colon carcinoma cells. PLK1 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing PLK1 ab32795 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of PLK1 using PLK1 Monoclonal antibody (13E8) ab32795 shows staining in HeLa cells. PLK1 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing PLK1 ab32795 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of PLK1 using PLK1 Monoclonal antibody (13E8) ab32795 shows staining in U251 glioma cells. PLK1 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing PLK1 ab32795 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry was performed on biopsies of deparaffinized Human testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing TPX2 ab32795 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • All lanes : Anti-TPX2 antibody [18D5-1] (ab32795)

    Lane 1 : MCF7 cells
    Lane 2 : MCF7 cells overexpressing Aurora A
    Lane 3 : MCF7 cells overexpressing TPX2

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated donkey anti-mouse IgG
    Developed using the ECL technique

    Predicted band size : 86 kDa

    Image from Grover A et al.,PLoS One. 2012;7(1):e30890. Epub 2012 Jan 27. Fig 7.; doi:10.1371/journal.pone.0030890; January 27, 2012, PLoS ONE 7(1): e30890.

Anti-TPX2 antibody [18D5-1] (ab32795) 使用論文

This product has been referenced in:
  • Platani M  et al. Mio depletion links mTOR regulation to Aurora A and Plk1 activation at mitotic centrosomes. J Cell Biol 210:45-62 (2015). ICC/IF ; Human . Read more (PubMed: 26124292) »
  • Wu JM  et al. Aurora kinase inhibitors reveal mechanisms of HURP in nucleation of centrosomal and kinetochore microtubules. Proc Natl Acad Sci U S A 110:E1779-87 (2013). Read more (PubMed: 23610398) »

See all 8 Publications for this product

Product Wall

OK, thanks. If it has never worked, with or without boiling the samples, and using the conditions you sent earlier, I think it should be replaced. For a replacement, we do not have other antibodies that have been validated for reactivity with mouse...

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Thank you for sending the details of your sample preparation. Have you stained blots of these samples for any other proteins? I think the preparation is good, though we recommend boiling samples to ensure complete denaturation. It is possible that the ...

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Thank you for contacting Abcam about this TPX2 western blot issue and for sending the image of the blot. Can you please give me a few details about the samples and the protocol? In particular, we are interested in the species and the nature of the samp...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (Spindles in X. laevis cytoplasmic egg extract)
Specification Spindles in X. laevis cytoplasmic egg extract
Fixative Formaldehyde and Methanol
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Nov 17 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - other (Xenopus laevis and tropicalis cytoplasmic egg extr)
Loading amount 40 µg
Specification Xenopus laevis and tropicalis cytoplasmic egg extr
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Nov 17 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (fibroblasts)
Specification fibroblasts
Fixative Paraformaldehyde
Permeabilization Yes - Methanol, 30 min at -20°C
Blocking step FCS as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: RT°C
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Dr. Ioannis Gavvovidis

Verified customer

投稿 Dec 02 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"