Overview

  • Product name
    Anti-NUP98 antibody [2H10]
    See all NUP98 primary antibodies
  • Description
    Rat monoclonal [2H10] to NUP98
  • Host species
    Rat
  • Tested applications
    Suitable for: ELISA, WB, Dot blot, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
  • Immunogen

    Recombinant fragment corresponding to Human NUP98 aa 1-466.

  • Positive control
    • WB: HeLa nuclear lysate. Jurkat, HeLa, COS-7, NIH/3T3 and SH-SY5Y cell lysate. ICC/IF: HeLa and NIH/3T3 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab50610 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
ELISA Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 98 kDa.
Dot blot Use a concentration of 0.5 - 1 µg/ml.
ICC/IF 1/100 - 1/200. Customers have reported that Paraformaldehyde/Triton x-100 fixation provides better results, with sharp, regularly punctuate perinuclear signals. In MetOH fixed cells, the signal intensity can be somehwat lower and fuzzier and that single nucleoporin dots can be harder to distinguish around nuclear chromatin. Please see images below.
IP Use at an assay dependent concentration.

Target

  • Function
    Nup98 and Nup96 play a role in the bidirectional transport across the nucleoporin complex (NPC). The repeat domain in Nup98 has a direct role in the transport.
  • Involvement in disease
    Note=A chromosomal aberration involving NUP98 is found in a form of acute myeloid leukemia. Translocation t(7;11)(p15;p15) with HOXA9. Translocation t(11;17)(p15;p13) with PHF23.
    Note=A chromosomal aberration involving NUP98 is found in childhood acute myeloid leukemia. Translocation t(5;11)(q35;p15.5) with NSD1. Translocation t(8;11)(p11.2;p15) with WHSC1L1.
    Note=A chromosomal aberration involving NUP98 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with TOP1.
    Note=A chromosomal aberration involving NUP98 is found in a form of T-cell acute lymphoblastic leukemia (T-ALL). Translocation t(3;11)(q12.2;p15.4) with LNP1.
    Note=A chromosomal aberration involving NUP98 is associated with pediatric acute myeloid leukemia (AML) with intermediate characteristics between M2-M3 French-American-British (FAB) subtypes. Translocation t(9;11)(p22;p15) with PSIP1/LEDGF. The chimeric transcript is an in-frame fusion of NUP98 exon 8 to PSIP1/LEDGF exon 4.
  • Sequence similarities
    Belongs to the nucleoporin GLFG family.
    Contains 1 peptidase S59 domain.
  • Domain
    Contains G-L-F-G repeats.
  • Post-translational
    modifications
    Isoform 1 to isoform 4 are autoproteolytically cleaved to yield Nup98 and Nup96 or Nup98 only, respectively. Cleaved Nup98 is necessary for the targeting of Nup98 to the nuclear pore and the interaction with Nup96.
  • Cellular localization
    Nucleus > nuclear pore complex. Nucleus membrane. Nup96 is localized to the nucleoplasmic side of the nuclear pore complex, at or near the nucleoplasmic basket.
  • Information by UniProt
  • Database links
  • Alternative names
    • 96 kDa nucleoporin antibody
    • 98 kDa nucleoporin antibody
    • ADAR2 antibody
    • ADIR2 antibody
    • GLFG-repeat containing nucleoporin antibody
    • Nuclear pore complex protein Nup96 antibody
    • Nuclear pore complex protein Nup98 Nup96 antibody
    • Nucleoporin 98kD antibody
    • nucleoporin 98kDa antibody
    • Nucleoporin Nup96 antibody
    • Nucleoporin Nup98 antibody
    • NUP196 antibody
    • NUP96 antibody
    • Nup98 antibody
    • Nup98-Nup96 antibody
    • NUP98_HUMAN antibody
    see all

Images

  • Lentiviral vector-encoded shRNAs achieve efficient knock-down of human nucleoporins and have negligible cytotoxic or cytostatic effects.

    HeLa cells (4×106) were transduced with lentiviral vectors (MOI 50) encoding shRNAs specific for the indicated nucleoporins and used at 2 days p.t for Nup153 shRNA and 5 days p.t for all others.

    (Panel B) Subcellular localisation of nuclear pore components upon nucleoporin knock-down was tested by confocal fluorescence microscopy of LV- (Control) and LV-shRNA transduced cells using specific anti-Nup antibodies. Images were acquired on the same day with the same conditions and are representative of two independent experiments.

  • All lanes : Anti-NUP98 antibody [2H10] (ab50610) at 1 µg/ml

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) nuclear lysate
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 4 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
    Lane 5 : COS-7 (african green monkey kidney fibroblast-like cell line) cell lysate
    Lane 6 : NIH/3T3 (mouse embyro fibroblast cell line) cell lysate
    Lane 7 : P19 cell lysate
    Lane 8 : NRK cell lysate

    Secondary
    All lanes : Goat Anti-Mouse IgG-Peroxidase

    Developed using the ECL technique.
  • ab50610 (1/100) staining NUP98 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (green).

    Cells were fixed with paraformaldehyde/Triton X-100 [10 min in PTEMF buffer (20mM PIPES, 1mM MgCl2, 10mM EGTA, 4% PFA) /0.2% Triton X-100 at room temperature] or methanol (6 min in methanol -20 °C , followed by 3 washes in 1x PBS) and counterstained with DAPI in order to highlight the nucleus (blue).

  • ab50610 (1/100) staining NUP98 in NIH/3T3 (Mouse embryo fibroblast cell line) cells (green).

    Cells were fixed with paraformaldehyde/Triton X-100 (10 min in PTEMF buffer (20mM PIPES, 1mM MgCl2, 10mM EGTA, 4% PFA) /0.2% Triton X-100 at room temperature) or methanol (6 min in methanol -20 °C , followed by 3 washes in 1x PBS) and counterstained with DAPI in order to highlight the nucleus (blue).

  • Paraformaldehyde-fixed, 0.5% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells stained for NUP98 (green) using ab50610 at 1/200 dilution in ICC/IF, followed by Donkey Anti-Rat Alexa Fluor® 488.

    See Abreview

References

This product has been referenced in:
  • Balmus G  et al. Targeting of NAT10 enhances healthspan in a mouse model of human accelerated aging syndrome. Nat Commun 9:1700 (2018). Read more (PubMed: 29703891) »
  • Lång A  et al. Visualization of PML nuclear import complexes reveals FG-repeat nucleoporins at cargo retrieval sites. Nucleus 8:404-420 (2017). Read more (PubMed: 28402725) »
See all 12 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Huh-7.5 (human hepatoma cells))
Permeabilization
No
Specification
Huh-7.5 (human hepatoma cells)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
PFA + glutaraldehyde

Abcam user community

Verified customer

Submitted Mar 15 2016

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Mouse Cell lysate - nuclear (stem cells)
Specification
stem cells
Total protein in input
30 µg

Abcam user community

Verified customer

Submitted Oct 13 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 18°C
Sample
Human Cell (293T)
Specification
293T
Permeabilization
Yes - 0.5% Triton X100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 14 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12 Bis tris)
Sample
Human Cell lysate - whole cell (293T)
Specification
293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

Abcam user community

Verified customer

Submitted May 14 2014

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

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