TPX2 Antibody - Azide and BSA Free

Novus Biologicals | Catalog # NB500-179

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Mammal

Cited:

Human, Mouse, Porcine, Bovine, Ovine

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Knockdown Validated

Cited:

Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Knockdown Validated

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

Azide and BSA Free
View all TPX2 Primary Antibodies »

Product Specifications

Immunogen

A recombinant segment of the C-terminal domain of human TPX2 [UniProt# Q9ULW0]

Localization

Nuclear. Staining of non-mitotic cells and cell cycle-dependent changes in abundance peaking at G2/M phase.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

100 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for TPX2 Antibody - Azide and BSA Free

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179]

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179]

Immunocytochemistry/Immunofluorescence: TPX2 Antibody [NB500-179] - Analysis of TPX2 at the mitotic spindle microtubules and poles in a HeLa metaphase cell. ICC/IF image submitted by a verified customer review.
Flow Cytometry: TPX2 Antibody [NB500-179]

Flow Cytometry: TPX2 Antibody [NB500-179]

TPX2-Antibody-Flow-Cytometry-NB500-179-img0005.jpg
Western Blot: TPX2 Antibody [NB500-179]

Western Blot: TPX2 Antibody [NB500-179]

Western Blot: TPX2 Antibody [NB500-179] - Analysis of TPX2 expression in 1) HeLa, 2) Ntera2, 3) K-562 and 4) Raji whole cell lysates using NB500-179.
Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179]

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179]

Immunocytochemistry/Immunofluorescence: TPX2 Antibody [NB500-179] - Staining of HeLa cells fixed in 3.5% paraformaldehyde using NB 500-179 (1:1,000). Nuclear staining during interphase and spindle staining during mitosis.
Simple Western: TPX2 Antibody [NB500-179]

Simple Western: TPX2 Antibody [NB500-179]

Simple Western: TPX2 Antibody [NB500-179] - Lane view shows a specific band for TPX2 using HeLa cell lysate and antibody at 1:250. Electropherogram image of corresponding Simple Western lane view. Image reported by internal validation.
TPX2 Antibody

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179] -

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179] - Depletion of TPX2 or Aurora-A reduces cell viability of BRCA2-deficient breast cancer cells. a HCC1806-shBRCA2dox, HCC38-shBRCA2dox, SUM149-shBRCA2dox, & MB231-shBRCA2dox were grown on coverslips & treated with doxycycline (3 days) and/or irradiated (IR, 5 Gy) as indicated. Subsequently, cells were stained for RAD51 & gamma H2AX. Scale bars represent 5 μm. b Quantification of results from a. Percentages of cells with ≥5 RAD51 foci per nucleus are indicated (n ≥ 31). c Percentages of cell survival of doxycycline-treated cells vs untreated cells, transfected with indicated siRNAs. Unpaired two-tailed t tests were used to test for statistical significance (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). d BT-549 cells were transfected with siTPX2 or control siRNA (CTR). Cells were grown on coverslips for 3 days after which they were incubated with EdU conjugated to azide-Alexa 488 (10 μM) for 15 min. Subsequently, cells were fixed & stained for 53BP1 & gamma H2AX. Amounts of 53BP1 & gamma H2AX foci per cell of at least 30 EdU-positive cells were counted. Means & standard deviations are depicted. Mann–Whitney U tests were used to analyze statistical significance (*p ≤ 0.05, ** = p ≤ 0.01, ***p ≤ 0.001, ns not significant). e BT-549 cells were transfected as in d, irradiated (IR, 5 Gy), & fixated 0.5 or 6 h after irradiation. Amounts of 53BP1 & gamma H2AX foci per cell were counted. Means & standard deviations are depicted. Mann–Whitney U tests were used to analyze statistical significance (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ns = not significant) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30177840), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TPX2 Antibody

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179] -

Immunocytochemistry/ Immunofluorescence: TPX2 Antibody [NB500-179] - TPX2 depletion preferentially affects cell viability in BRCA2-deficient cancer cells. a BT-549-shBRCA2dox cells were left untreated or were treated with doxycycline (2 or 4 days), & subsequently harvested for western blotting for BRCA2 & actin. b BT-549-shBRCA2dox cells were treated as in panel A, & mRNA expression levels of BRCA2 were analyzed relative to GAPDH using qRT-PCR. c BT-549-shBRCA2dox cells were grown on coverslips & treated with doxycycline (3 days) and/or irradiated (IR, 5 Gy) as indicated. At 3 h after irradiation, cells were fixed & analyzed for RAD51 & gamma H2AX foci formation. Scale bars represent 5 μm. d Percentages of cells with ≥5 RAD51 foci per nucleus are indicated. (n ≥ 50 per condition). e BT-549-shBRCA2dox cells were treated with doxycycline (3 days) & were subsequently transfected with indicated siRNAs. A total of 30,000 cells were plated 48 h following transfection. Viable cells were counted 5 days later. Percentages of cell survival of doxycycline-treated vs untreated cells are depicted. Error bars indicate standard deviations of two experimental replicates. Unpaired two-tailed t tests were used to test for statistical significance (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30177840), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TPX2 Antibody - Azide and BSA Free

Western Blot: TPX2 Antibody - Azide and BSA Free [NB500-179] -

CtIP depletion causes improper progression of mitosis. (A) The progression of mitosis in HeLa cells was monitored by time-lapse microscopy. HeLa cells were transfected with control or CtIP-1 or siRNA. After 48 h, control and CtIP-depleted HeLa cells were seeded in 12-well plates and transfected with GFP-tagged histone H2B. Fluorescent images were obtained every 5 min starting at the stage of nuclear envelope breakdown. (B) A quantification of the time from nuclear envelope breakdown to anaphase onset in control cells and CtIP-depleted cells. Bars represent the mean +/- SD from three independent experiments. **, p < 0.01, compared to control cells. (C) Delayed mitosis progression in CtIP-depleted cells was confirmed by prolonged phosphorylation of histone H3 (pH3S10). Control and CtIP-depleted HeLa cells were synchronized with a double thymidine block to arrest at the G1/S boundary and released from this block for indicated times. Total proteins collected at the indicated times after release were analyzed by Western blotting using anti-pH3S10 antibody. Histone H3 antibody was used as a loading control. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/2073-4409/11/18/2814), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TPX2 Antibody - Azide and BSA Free

Western Blot: TPX2 Antibody - Azide and BSA Free [NB500-179] -

CtIP interacts with TPX2. (A,B) CtIP coimmunoprecipitates with TPX2. Total cell lysates (1 mg) from HEK293T cells transfected with Flag-tagged full length CtIP and HA-tagged full length TPX2 were immunoprecipitated with anti-Flag (A) or anti-HA (B) antibodies. Immunoblotting was then performed with the indicated antibodies. (C) Total cell lysates from HEK293T cells transfected with control siRNA and CtIP siRNA were immunoprecipitated with anti-CtIP or anti-TPX2 antibodies. Immunoblotting was then performed with the indicated antibodies. (D) The cellular localization of CtIP and TPX2 during mitosis was monitored using Immunofluorescence microcopy. Asynchronous HeLa cells were fixed and stained with anti-CtIP and anti-TPX2 antibodies. Representative images show that CtIP colocalizes with TPX2 at the kinetochore from prometaphase through anaphase. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/2073-4409/11/18/2814), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TPX2 Antibody - Azide and BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 24019927)

Immunocytochemistry/ Immunofluorescence

1:1000

Immunoprecipitation

1:10-1:100

Knockdown Validated

reported in scientific literature (PMID 31586073)

Simple Western

1:250

Western Blot

1:1000
Application Notes

In WB a band at approx. 100 kDa can be detected. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.2 mg/mL, separated by Size, antibody dilution of 1:250, apparent MW was 89 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 5 using NB500-179 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Whole antisera

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: TPX2

TPX2 (targeting protein for XKLP2) is an ortholog of the Targeting Protein for Xenopus kinesin-like protein 2 (XKLP2) and was originally named restricted expression/proliferation-associated protein 100 (REPP86). TPX2 regulates the Xenopus kinesin-like protein 2 which is a plus end-directed microtubule motor required for centrosome separation and maintenance of spindle polarity during mitosis. TPX2 levels steadily increase during cell cycle progression with highest expression levels in M phase and lowest levels in G1 phase. TPX2 activates mitotic serine-threonine Aurora A kinase, nucleates microtubules and is considered as a critical factor for spindle assembly in mammalian mitotic cells. It is actively transported into the nucleus during interphase via importin alpha/beta receptors and this nuclear import sequesters TPX2 away from cytoplasmic tubulin to prevent premature spindle assembly. TPX2 haploinsufficiency, leading to decreased levels of TPX2, significantly increases the susceptibility for tumor development in mice and cells deficient in the Aurora A/TPX2 complex presents short spindles which, despite of being able to segregate chromosomes, result in mitotic failure.

Alternate Names

C20orf1HCTP4, C20orf2, chromosome 20 open reading frame 1, Differentially expressed in cancerous and non-cancerous lung cells 2, differentially expressed in lung cells, DIL2, DIL-2FLS353, HCA519, Hepatocellular carcinoma-associated antigen 519, P100, p100GD:C20orf1, preferentially expressed in colorectal cancer, Protein fls353, REPP86, restricted expression proliferation associated protein 100, Restricted expression proliferation-associated protein 100, targeting protein for Xklp2, TPX2, microtubule-associated protein homolog, TPX2, microtubule-associated, homolog (Xenopus laevis)

Entrez Gene IDs

22974 (Human); 72119 (Mouse); 311546 (Rat)

Gene Symbol

TPX2

UniProt

Q9ULW0 (Human)

Additional TPX2 Products

Product Documents for TPX2 Antibody - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for TPX2 Antibody - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for TPX2 Antibody - Azide and BSA Free

Customer Reviews for TPX2 Antibody - Azide and BSA Free (1)

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Customer Images

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  • Name: Patrizia Lavia
    Application: Immunofluorescence
    Sample Tested: See PDF
    Species: Human
    Verified Customer | Posted 01/16/2015
    TPX2 at the mitotic spindle microtubules and poles in a HeLa metaphase cell.
    TPX2 Antibody - Azide and BSA Free NB500-179

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Protocols

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FAQs for TPX2 Antibody - Azide and BSA Free

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  • Q: In what assays cross-reactivity of NB500-179 with mouse has been confirmed? Do you have any information, or are there by chance any mouse images that just haven't made it onto the datasheet yet?

    A: We tested this ab on mouse cells in a Western blot with good results.

  • Q: We know that the immunogen is a recombinant segment of the C-terminal domain of human TPX2. If it is possible, we would be interested to know the exact epitope within this domain.

    A: Unfortunately, the exact immunogen sequence for NB500-179 is considered proprietary. It comes from the last 100 amino acids of TPX2.

  • Q: In what assays cross-reactivity of NB500-179 with mouse has been confirmed? Do you have any information, or are there by chance any mouse images that just haven't made it onto the datasheet yet?

    A: We tested this ab on mouse cells in a Western blot with good results.

  • Q: We know that the immunogen is a recombinant segment of the C-terminal domain of human TPX2. If it is possible, we would be interested to know the exact epitope within this domain.

    A: Unfortunately, the exact immunogen sequence for NB500-179 is considered proprietary. It comes from the last 100 amino acids of TPX2.

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