Key features and details
- Mouse monoclonal [18D5-1] to TPX2
- Suitable for: ICC, WB, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Product nameAnti-TPX2 antibody [18D5-1]
See all TPX2 primary antibodies
DescriptionMouse monoclonal [18D5-1] to TPX2
SpecificityThis antibody detects TPX2.
Tested applicationsSuitable for: ICC, WB, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Human
Recombinant TPX2 protein (Human).
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
Concentration information loading...
PurityProtein G purified
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.
|ICC||Use at an assay dependent concentration.|
|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.|
FunctionSpindle 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 specificityExpressed in lung carcinoma cell lines but not in normal lung tissues.
Sequence similaritiesBelongs to the TPX2 family.
Developmental stageExclusively expressed in proliferating cells from the transition G1/S until the end of cytokinesis.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationNucleus. 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
- C20ORF1 antibody
- C20orf2 antibody
- Chromosome 20 Open Reading Frame 1 antibody
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.
All lanes : HRP-conjugated donkey anti-mouse IgG
Developed using the ECL technique.
Predicted band size: 86 kDa
ab32795 has been referenced in 21 publications.
- Zhao L et al. Identification of biomarkers for the transition from low-grade glioma to secondary glioblastoma by an integrated bioinformatic analysis. Am J Transl Res 12:1222-1238 (2020). PubMed: 32355537
- Huang DH et al. TPX2 silencing exerts anti-tumor effects on hepatocellular carcinoma by regulating the PI3K/AKT signaling pathway. Int J Mol Med 44:2113-2122 (2019). PubMed: 31638175
- Tian Y et al. TPX2 gene silencing inhibits cell proliferation and promotes apoptosis through negative regulation of AKT signaling pathway in ovarian cancer. J Cell Biochem 119:7540-7555 (2018). PubMed: 29904936
- Chen M et al. Targeting TPX2 suppresses proliferation and promotes apoptosis via repression of the PI3k/AKT/P21 signaling pathway and activation of p53 pathway in breast cancer. Biochem Biophys Res Commun 507:74-82 (2018). PubMed: 30454896
- Hsu WH et al. Adducin-1 is essential for spindle pole integrity through its interaction with TPX2. EMBO Rep 19:N/A (2018). PubMed: 29925526