Anti-Nuclear Pore Complex Proteins antibody [Mab414] - ChIP Grade (ab24609)

Overview

  • Product name
    Anti-Nuclear Pore Complex Proteins antibody [Mab414] - ChIP Grade
    See all Nuclear Pore Complex Proteins primary antibodies
  • Description
    Mouse monoclonal [Mab414] to Nuclear Pore Complex Proteins - ChIP Grade
  • Specificity
    ab24609 reacts with Nuclear Pore Complex (NPC) Proteins. Aris et al ( J. Cell Biol. 108:2059-2067, 1989) report "In rat liver, this monoclonal antibody, mAb 414, binds to nuclear pore complex proteins, including one of molecular weight 62,000 (Davis, L. I., and G. Blobel. 1987. Proc. Natl. Acad. Sci. USA. 84:7552-7556). In yeast, mAb 414 cross reacts by immunoblotting with two proteins that have apparent molecular weights of 110,000 and 95,000, and are termed p110 and p95, respectively. Examination of subcellular fractions by immunoblotting shows that both p110 and p95 are located exclusively in the nuclear fraction. The mAb 414 immunoprecipitates several proteins from a crude yeast cell extract, including p110, p95, and a approximately 55-kD protein. Immunoprecipitation from subcellular fractions yields only p110 and p95 from purified nuclei, whereas the approximately 55-kD protein is immunoprecipitated from the soluble fraction. Digestion of purified nuclei with DNase to produce nuclear envelopes releases some of p110, but the majority of p110 is solubilized only after treatment of envelopes with 1 M NaCl. Immunofluorescence localization using yeast cells and isolated nuclei shows a punctate and patchy staining pattern of the nucleus."
  • Tested applications
    Suitable for: ChIP, WB, ICC/IF, IP, Electron Microscopy, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cat, Human, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish
    Predicted to work with: Vertebrata
  • Immunogen

    Nuclear pore complex mixture.

  • Epitope
    ab24609 recognizes the conserved domain FXFG repeats in nucleaporins like the p62, p152, p90.
  • Positive control
    • Raw, HEK 293 cell lysate (see Abreview), rat liver lysate (see Aris reference)
  • General notes


    This is a reliable general purpose monoclonal antibody which recognizes a related family of NPC proteins. This antibody is ideal for studying the morphology and composition of the nucleus and nuclear envelope. It is also useful in studying changes in the nuclear structure during mitosis and meiosis.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.03% Thimerosal (merthiolate)
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    This is a reliable general purpose monoclonal antibody which recognizes a related family of NPC proteins. This antibody is ideal for studying the morphology and composition of the nucleus and nuclear envelope. It is also useful in studying changes in the nuclear structure during mitosis and meiosis.
  • Clonality
    Monoclonal
  • Clone number
    Mab414
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab24609 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
ChIP Use at an assay dependent concentration. PubMed: 20419146
WB 1/5000. Predicted molecular weight: 62 kDa. Suitable also in non reduced western blotting conditions (see Abreview). Nuclear extraction may be necessary (protocol detailed in Aris et al, http://www.jcb.org/cgi/reprint/108/6/2059).
ICC/IF 1/5000. Fix cells with 4% paraformaldehyde in NWB (200 mM sucrose, 15 mM Hepes, pH 7.4, 50 mM NaCl, 2.5 mM MgCl2, and 1 mM DTT). Permeabilise with 0.1% NP-40 or 0.1% Triton X-100 in PBS for 2 min. (see Lopez-Soler reference); different customer have used this antibody at different dilutions for ICC/IF (see images below). We recommend that optimal working dilutions are determined by each customer.
IP 1/5000. Nuclear extraction may be necessary (protocol detailed in Aris et al, http://www.jcb.org/cgi/reprint/108/6/2059).
Electron Microscopy 1/5000.
IHC-Fr Use at an assay dependent concentration. 4% paraformaldehyde fixed tissue cut on a cryostat (see Kimura reference).

Target

Images

  • ab24609 staining nuclear pore complex proteins in Human chronic myelogenous leukemia cells from bone marrow cells. Cells were fixed with 4% paraformaldehyde in DPBS buffer for 20 mins and permeabilised with 0.1% Triton X-100 for 20 min at room temperature. Cells were blocked with 2% normal goat serum in DPBS with 1% BSA and washed with 0.1% Tween 20.

  • ab24609 (1/200) staininig nuclear pore complex proteins in human RPE-1 cells (green). Cells were fixed in paraformaldehyde, permeabilised with Triton X100 and counterstained with DAPI in order to highlight the nucleus (blue). Please refer to abreview for further experimental details.

    See Abreview

  • ab24609 (1/500) staining nuclear pore complex proteins in human Hela 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-X100 at room T°C) or Methanol (6 min in Methanol -20 °C , followed by 3 washes in 1x PBS) and counterstained with Dapi in order to highligh the nucleus (blue).
  • ab24609 (1/500) staining nuclear pore complex proteins in murine NIH/3T3 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-X100 at room T°C) or Methanol (6 min in Methanol -20 °C , followed by 3 washes in 1x PBS) and counterstained with Dapi in order to highligh the nucleus (blue).

References

This product has been referenced in:
  • Capitanio JS  et al. Human Nup98 regulates the localization and activity of DExH/D-box helicase DHX9. Elife 6:N/A (2017). Read more (PubMed: 28221134) »
  • Kinosada H  et al. HTLV-1 bZIP Factor Enhances T-Cell Proliferation by Impeding the Suppressive Signaling of Co-inhibitory Receptors. PLoS Pathog 13:e1006120 (2017). IF ; Mouse . Read more (PubMed: 28046066) »

See all 53 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (DLD1 Cells)
Permeabilization
Yes - 0.5% TritonX-100
Specification
DLD1 Cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 24 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Fruit fly (Drosophila melanogaster) Cell (OSS cells)
Specification
OSS cells
Permeabilization
Yes - Tween 0,2%
Fixative
Formaldehyde
Username

Ms. Nathalie Gred Gueguen

Verified customer

Submitted Nov 12 2014

Application
IHC - Wholemount
Sample
Fruit fly (Drosophila melanogaster) Tissue (Imaginal Wing Disc)
Specification
Imaginal Wing Disc
Username

Peng

Verified customer

Submitted Jul 09 2014

The staining pattern of this antibody should be nuclear or nuclear membrane.

Western blot

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - nuclear (U2OS)
Specification
U2OS
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2Os)
Specification
U2Os
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton in PBS
Blocking step
FCS in PBS as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Ms. Anna Löschberger

Verified customer

Submitted Oct 15 2012

Thank you for contacting us.



While this product is predicted to react with all vertebrates, we have not yet tested this with cow. For UNTESTED species and/or applications, we have established a testing discount program. Here is a...

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I am very pleased to hear you would like to accept our offer and test one or both of these products in blocking experiements. Each code is associated with a paricular antibody so be sure tho reference the correct code when ordering and submitting the A...

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Thank you very much for your interest in our Nuclear Pore Complex Proteins antibodies. I have spoken to the labs about these product and have found that these are indeed the same clone detailed inin the papers by Davis & Blobel as we spoke of. Read More

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