Anti-Nup153 antibody [QE5] (ab24700)

Mouse monoclonal Nup153 antibody [QE5]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Human. Cited in 34 publication(s). Independently reviewed in 4 review(s).

Overview

  • Product name
    Anti-Nup153 antibody [QE5]
    See all Nup153 primary antibodies
  • Description
    Mouse monoclonal [QE5] to Nup153
  • Host species
    Mouse
  • Specificity
    ab24700 recognises NUP153 as well as two related nuclear pore complex proteins: NUP214 and p62.
  • Tested applications
    Suitable for: Flow Cyt, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Full length protein: rat liver nuclear envelope proteins

  • Positive control
    • In Immunocytochemistry, this antibody gave a positive signal in HepG2 cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab24700 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 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB 1/1000. Detects a band of approximately 154 kDa (predicted molecular weight: 154 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 23423481

Target

  • Function
    Possible DNA-binding subunit of the nuclear pore complex (NPC). The repeat-containing domain may be involved in anchoring components of the pore complex to the pore membrane.
  • Sequence similarities
    Contains 4 RanBP2-type zinc fingers.
  • Domain
    Contains F-X-F-G repeats.
  • Cellular localization
    Nucleus > nuclear pore complex. Located to the terminal ring structure of the nucleoplasmic cage.
  • Information by UniProt
  • Database links
  • Alternative names
    • 153 kDa nucleoporin antibody
    • HNUP153 antibody
    • N153 antibody
    • NU153_HUMAN antibody
    • Nuclear pore complex protein hnup153 antibody
    • Nuclear pore complex protein Nup153 antibody
    • Nucleoporin 153kDa antibody
    • Nucleoporin Nup153 antibody
    • Nup 153 antibody
    • Nup153 antibody
    see all

Images

  • Immunocytochemical analysis of U2OS cells, labeling Nup153 with ab24700. Cells were formaldehyde fixed, permeabilized with NP40, and blocked with 3% BSA for 1 hour at 21°C. Immunostaining with ab24700 diluted 1/100 for 12 hours at 4°C.

    See Abreview

  • Anti-Nup153 antibody [QE5] (ab24700) at 1/1000 dilution + Human U2OS nuclear lysate at 40 µg

    Secondary
    HRP-conjugated donkey anti-mouse IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 154 kDa
    Observed band size: 160 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 minutes

    See Abreview

  • Human and mouse cells treated with different concentrations of ab24700. The cells were fixed in 4% formaldehyde and permeabilized in 0.2% Triton x100 for 10 minutes at room temperature, then washed 3 times in PBS. The fixation and permeabilization was carried out in a single step. The image shows clear staining of the nuclear envalope (green). The DNA is stained with DAPI (blue).

    A: HeLa cells, ab24700 used at 4µg/ml
    B: 3T3 cells, ab24700 used at 2µg/ml
    C: 3T3 cells, ab24700 used at 4µg/ml
    D: 3T3 cells, ab24700 used at 6.6µg/ml

  • Overlay histogram showing HeLa cells stained with ab24700 (red line). The cells were fixed with 80% methanol (5 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 (ab24700, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ICC/IF image of ab24700 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24700, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • Methanol fixed HeLa stained with ab24700. This antibody brilliantly highlights the nuclear membrane (green). The golgi is stained with Giantin (yellow).
  • Human and mouse cells treated with different concentrations of ab24700. The cells were fixed in 100% methanol for 20 minutes at -20°C, then washed once in PBS. The image shows clear staining of the nuclear envalope (green). The DNA is stained with DAPI (blue).

    A: HeLa cells, ab24700 used at 4µg/ml
    B: 3T3 cells, ab24700 used at 2µg/ml
    C: 3T3 cells, ab24700 used at 4µg/ml
    D: 3T3 cells, ab24700 used at 6.6µg/ml

References

This product has been referenced in:
  • Vuorinen EM  et al. Depletion of nuclear import protein karyopherin alpha 7 (KPNA7) induces mitotic defects and deformation of nuclei in cancer cells. BMC Cancer 18:325 (2018). Read more (PubMed: 29580221) »
  • Re A  et al. Nucleoporin 153 regulates estrogen-dependent nuclear translocation of endothelial nitric oxide synthase and estrogen receptor beta in prostate cancer. Oncotarget 9:27985-27997 (2018). Read more (PubMed: 29963256) »
See all 34 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (SAOS-2 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
SAOS-2 cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Dec 18 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - 0.5% Triton
Specification
U2OS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 27 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - NP40
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 15 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - nuclear (U2OS)
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Apr 22 2014

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

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