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

Overview

  • Product nameAnti-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
  • Specificityab24609 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 applicationsSuitable 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: all other Vertebrates
  • Immunogen

    Nuclear pore complex mixture.

  • Epitopeab24609 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

  • FormLiquid
  • Storage instructionsShipped 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 bufferPreservative: 0.03% Thimerosal (merthiolate)
    Constituent: PBS
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Primary antibody notesThis 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.
  • ClonalityMonoclonal
  • Clone numberMab414
  • IsotypeIgG1
  • 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

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

This product has been referenced in:
  • Cundell MJ  et al. A PP2A-B55 recognition signal controls substrate dephosphorylation kinetics during mitotic exit. J Cell Biol 214:539-54 (2016). Read more (PubMed: 27551054) »
  • Guan Y  et al. Phosphorylation of Def Regulates Nucleolar p53 Turnover and Cell Cycle Progression through Def Recruitment of Calpain3. PLoS Biol 14:e1002555 (2016). WB ; Zebrafish . Read more (PubMed: 27657329) »

See all 31 Publications for this product

Product Wall

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

Thank you very much for your interest in ab24609.

To our knowledge, ab24609 has not been tested in IHC-P. Therefore, I can offer a discount off a future purchase if you buy ab24609 now, test it inIHC-P and submit feedback to us in the form o...

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