Anti-Nup153 antibody [QE5] (ab24700)

Reviews (4)Q&A (2)References (34)

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

    • Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)This image is courtesy of an anonymous Abreview.

      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

    • Western blot - Anti-Nup153 antibody [QE5] (ab24700)
      Western blot - Anti-Nup153 antibody [QE5] (ab24700)This image is courtesy of an anonymous 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

    • Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)This image is courtesy of Patrizia Lavia & Maria de Luca, University La Sapienza

      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

    • Flow Cytometry - Anti-Nup153 antibody [QE5] (ab24700)
      Flow Cytometry - Anti-Nup153 antibody [QE5] (ab24700)
      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      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.
    • Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      Methanol fixed HeLa stained with ab24700. This antibody brilliantly highlights the nuclear membrane (green). The golgi is stained with Giantin (yellow).
    • Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)
      Immunocytochemistry/ Immunofluorescence - Anti-Nup153 antibody [QE5] (ab24700)This image is courtesy of Patrizia Lavia & Maria de Luca, University La Sapienza

      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

    Publishing research using ab24700? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-Nup153 antibody [QE5]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 18 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Nup153 antibody [QE5]

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 27 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Nup153 antibody [QE5]

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 15 2015

    Western blot abreview for Anti-Nup153 [QE5] antibody

     Excellent
    Abreviews
    abreview image
    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%
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 22 2014

    Question

    For your QE5 Nup153 antibody, what is the exact epitope the ab has been raised against (N, Zn finger, or C term)?

    Read More
    Answer

    Thank you for your enquiry. The antibody was raised against rat liver nuclear envelope proteins extracted in TX buffer (50 mM triethanolamine [TEA], pH 7.4, 500 mM NaCI, 0.5 % Triton X-100, 1 mM DTT, 1 mM PMSF and 1:1,000 CLAP [10 mg/ml each of chymostatin, leupeptin, antipain, and pepstatin in DMSO]). The exact epitope has been mapped to a region lying between residues 1,301 and 1,319. I enclude below the original paper that designed this clone and identified that epitope. http://www.jcb.org/cgi/reprint/126/3/603 Pante N et al. Interactions and three-dimensional localization of a group of nuclear pore complex proteins. J Cell Biol 126:603-17 (1994). PubMed: 8045926 I hope this information helps. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    I was wondering if you could show a western blot using your nup153 antibody so I can see how well it picks up the nup153 protein and how clean the antibody is. Thanks

    Read More
    Answer

    Thank you for your enquiry. I have been in touch with the source of this antibody and unfortunately I do not have a western image available at this time. I do however have soem images used to QC ths image by immunofluorescence. I have sent a follow up email with an image of the staining using this antibody. There is clear labeling of the nuclear lamina. A secondary alone control is used as a negative control. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

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