Overview

  • Product nameAnti-TNPO3 antibody [3152C2a]
    See all TNPO3 primary antibodies
  • Description
    Mouse monoclonal [3152C2a] to TNPO3
  • Tested applicationsSuitable for: Flow Cyt, WB, Dot blot, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment (Human) from near the N terminus.

  • Positive control
    • HeLa whole cell lysate; NIH3T3 whole cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab54353 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 Use 1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
WB 1/50 - 1/100. Detects a band of approximately 90 kDa (predicted molecular weight: 110 kDa).
Dot blot Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/300.

Target

Anti-TNPO3 antibody [3152C2a] images

  • Anti-TNPO3 antibody [3152C2a] (ab54353) at 1/100 dilution + HeLa whole cell lysate at 25 µg

    Secondary
    anti Mouse IgG at 1/2500 dilution

    Predicted band size : 110 kDa
    Observed band size : ~90 kDa (why is the actual band size different from the predicted?)
  • Anti-TNPO3 antibody [3152C2a] (ab54353) at 1/50 dilution + NIH3T3 whole cell lysate at 25 µg

    Secondary
    anti Mouse IgG at 1/2500 dilution

    Predicted band size : 110 kDa
    Observed band size : ~90 kDa (why is the actual band size different from the predicted?)
  • IHC image of ab54353 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab54353, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Overlay histogram showing HeLa cells stained with ab54353 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54353, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.

References for Anti-TNPO3 antibody [3152C2a] (ab54353)

This product has been referenced in:
  • Zehorai E & Seger R Beta-like importins mediate the nuclear translocation of mitogen-activated protein kinases. Mol Cell Biol 34:259-70 (2014). Read more (PubMed: 24216760) »
  • Melià MJ  et al. Limb-girdle muscular dystrophy 1F is caused by a microdeletion in the transportin 3 gene. Brain 136:1508-17 (2013). Read more (PubMed: 23543484) »

See all 9 Publications for this product

Product Wall

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Based on the details you provided, I believe this antibody is not working as we would expect. Although the product has not been specifically validated for frozen sections, I would e...

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I am sorry that you have been experiencing difficulties with this antibody in IHC. In order to assist you further, I was hoping you would be able to provide some additional information reg...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (ES cells)
Loading amount 100000 cells
Specification ES cells
Gel Running Conditions Non-reduced Denaturing (NuPage 4-12% Bis-Tris)
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

Submitted Apr 23 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela cells)
Specification Hela cells
Fixative Methanol
Permeabilization No
Blocking step Fish scale gelatin in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 11 2010

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