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

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Overview

Product name

Anti-Nuclear Pore Complex Proteins antibody [Mab414] - ChIP Grade
See all Nuclear Pore Complex Proteins products (4) ...

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

ChIP, WB, ICC/IF, IP, Electron Microscopy, IHC-Frmore details

Cross reactivity

Reacts with

Mouse, Rat, Cat, Human, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Zebrafish

Predicted to work with

all other Vertebrates

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)

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.03% Thimerosal (merthiolate)
Constituents: PBS

Concentration

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

Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Tags & Cell Markers >> Subcellular Markers >> Nucleus >> Nuclear Pore

  • Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609) image (enlarge)

Applications

Show applications key

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.

  • ShowHide

    ChIP

     ChIP: Use at an assay de...Read more →

    ChIP: Use at an assay dependent dilution. (PubMed: 20419146)

  • ShowHide

    WB

    Four stars (1 Abreview) WB: 1/5000Predicted mole...Read more →

    WB: 1/5000Predicted 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).)

  • ShowHide4 Images

    ICC/IF

    Five stars (2 Abreviews) ICC/IF: 1/5000(Fix cells...Read more →

    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.)

  • ShowHide

    IP

     IP: 1/5000(Nuclear extra...Read more →

    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

     EM: 1/5000

    EM: 1/5000

  • ShowHide

    IHC-Fr

     IHC-Fr: Use at an assay ...Read more →

    IHC-Fr: Use at an assay dependent dilution. (4% paraformaldehyde fixed tissue cut on a cryostat (see Kimura reference).)

Target

Relevance

The Nuclear pore complex (NPC) has a molecular mass of ~125 KDa in vertebrates and contains about 50 or more different proteins . The NPC spans the dual membrane of the the nuclear envelope (NE) and acts as a gateway for macromolecular traffic between the cytoplasm and the nucleus. The basic framework of the NPC consists of a central core with a ring-spoke structure exhibiting 8 fold radial symmetry. From this central ring 50 to 100 nm fibrils extend into the nucleoplasm and the cytoplasm. The NPC is in turn anchored in the NE by the nuclear lamina, a meshwork of lamins and lamin-associated proteins that forms a 15 nm thick fibrous structure between the inner nuclear membrane and peripheral chromatin. A number of proteins called nucleoporins have been localised to discrete regions of the NPC and are often used as markers for this compartment, e.g. Nup153. Approximately half of the nucleoporins (or Nups) contain a phenyalanine-glycine repeat motif (FG repeat), which may be diagnostic for proteins playing a role in nuclear transport.

Cellular localization

Nuclear

Alternative names

  • 107 kDa nucleoporin antibody
  • NPC proteins antibody
  • Nuclear pore complex protein Nup107 antibody
  • Nucleoporin 107kDa antibody
  • Nucleoporin Nup107 antibody
  • NUP 107 antibody
  • NUP 84 antibody
  • NUP107 antibody
  • NUP153 antibody
  • NUP155 antibody
  • NUP84 antibody
  • NUP98 antibody
see all

Database links

see all

Anti-Nuclear Pore Complex Proteins antibody [Mab414] - ChIP Grade images:

  Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)

Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)

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.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

See Abreview

  Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)

Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)

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).

Image and protocol kindly provided by Rosamaria Mangiacasale, Marilena Ciciarello and Patrizia Lavia, Univ Rome

  Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)

Immunocytochemistry/ Immunofluorescence - Nuclear Pore Complex Proteins antibody [Mab414] (ab24609)

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).

Image and protocol kindly provided by Rosamaria Mangiacasale, Marilena Ciciarello and Patrizia Lavia, Univ Rome

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

This product has been referenced in:

  • Beshore ELet al. C. elegans Dicer interacts with the P-granule component GLH-1 and both regulate germline RNPs. Dev Biol 350:370-81 (2011). ICC/IF; Caenorhabditis elegans.Read more (PubMed: 21146518) »
  • Chen Yet al. Functional k(v)10.1 channels localize to the inner nuclear membrane. PLoS One 6:e19257 (2011). ICC/IF.Read more (PubMed: 21559285) »

See all 13 publications for this product

Publishing research using ab24609? Please let us know so that we can cite the reference in this datasheet

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