Anti-NSP1 antibody [32D6] (ab4641)

Reviews (1)Q&A (1)References (7)
Immunocytochemistry/ Immunofluorescence - Anti-NSP1 antibody [32D6] (ab4641)

    Key features and details

    • Mouse monoclonal [32D6] to NSP1
    • Suitable for: ICC/IF, WB
    • Reacts with: Saccharomyces cerevisiae
    • Isotype: IgG

    Overview

    • Product name

      Anti-NSP1 antibody [32D6]

    • Description

      Mouse monoclonal [32D6] to NSP1

    • Host species

      Mouse

    • Specificity

      This antibody was initially identified as directed against a nuclear pore complex antigen by immunofluorescence localization. A screen of a lgt11 expression library yielded a single positive clone carrying an insert bearing ~66% of the C-terminal portion of NSP1. To confirm that 32D6 possessed high affinity for NSP1, strain RS453, which expresses a shortened isoform of NSP1, was compared to the wildtype strain BJ5465. NSP1 in RS453 contains an internal deletion that removes the coding sequence for 6 FXFG repeats, which are not essential for function and encodes a protein that has been observed to migrate at approxmately 85 kDa on SDS gels. The predicted size of wildtype NSP1 is 86.5 kDa, but NSP1 has been observed to migrate on SDS gels at ~100 kDa. In our gels, the apparent molecular mass of wildtype NSP1 was 108 kDa, whereas the short isoform of NSP1 in RS453 migrated at 91 kDa. The detection of two protein bands of apparent sizes 108 kDa and 91 kDa in BJ5465 and RS453, respectively, demonstrated that 32D6 recognized the pore complex protein NSP1.

    • Tested applications

      Suitable for: ICC/IF, WBmore details

    • Species reactivity

      Reacts with: Saccharomyces cerevisiae

    • Immunogen

      This was raised against a yeast nucleolar prep, and screened by immunocytochemistry and western blotting.

    • General notes


      This antibody is a useful marker of yeast nuclear pores.

      Previously labelled as Nsp1p.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    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: 0.065% Sodium azide
      Constituent: Tissue culture supernatant
    • Concentration information loading...

    • Purity

      Tissue culture supernatant

    • Purification notes

      Sterile filtered.

    • Primary antibody notes

      This antibody is a useful marker of yeast nuclear pores.

    • Clonality

      Monoclonal

    • Clone number

      32D6

    • Isotype

      IgG

    • Research areas

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab4641 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
    ICC/IF (1)
    1/1000.

    Yeast cells.
    WB
    1/10000 - 1/50000. Detects a band of approximately 108,91 kDa (predicted molecular weight: 85 kDa).

    1/10000 (cell lysates) - 1/50000 (nuclear fractions). For other (non-ECL) detection methods, 1/1000 - 1/5000.
    Notes
    ICC/IF
    1/1000.

    Yeast cells.
    WB
    1/10000 - 1/50000. Detects a band of approximately 108,91 kDa (predicted molecular weight: 85 kDa).

    1/10000 (cell lysates) - 1/50000 (nuclear fractions). For other (non-ECL) detection methods, 1/1000 - 1/5000.

    Target

    • Relevance

      NSP1 functions as a component of the nuclear pore complex (NPC). It plays an important role in several nuclear transport pathways including poly(A)+ RNA, tRNA, pre-ribosome, signal recognition particle (SRP), and protein transport.

    • Cellular localization

      Nuclear membrane: part of the nuclear pore complex, it presents a symmetric distribution with a cytoplasmic and a nucleoplasmic side.

    • Database links

      • Alternative names

        • NSP1 antibody
        • Nsp1p antibody
        • Nuclear pore protein NSP1 antibody
        • Nucleoporin NSP1 antibody
        • p110 antibody
        see all

      Images

      • Immunocytochemistry/ Immunofluorescence - Anti-NSP1 antibody [32D6] (ab4641)
        Immunocytochemistry/ Immunofluorescence - Anti-NSP1 antibody [32D6] (ab4641)Image courtesy of an anonymous Abreview.

        ab4641 (1/5000) detecting NSP1 in Saccharomyces cerevisae DDY1810 (green). Cells were fixed in paraformaldehyde, permeabilized in methanol/acetone and counterstained with DAPI in order to highlight the nucleus (blue). For further experimental details please refer to Abreview.

        See Abreview

      References (7)

      Publishing research using ab4641? Please let us know so that we can cite the reference in this datasheet.

      ab4641 has been referenced in 7 publications.

      • Bittmann J  et al. An advanced cell cycle tag toolbox reveals principles underlying temporal control of structure-selective nucleases. Elife 9:N/A (2020). PubMed: 32352375
      • Zhang W  et al. Brr6 and Brl1 locate to nuclear pore complex assembly sites to promote their biogenesis. J Cell Biol 217:877-894 (2018). PubMed: 29439116
      • Rüthnick D  et al. Characterization of spindle pole body duplication reveals a regulatory role for nuclear pore complexes. J Cell Biol 216:2425-2442 (2017). PubMed: 28659328
      • Colombi P  et al. The transmission of nuclear pore complexes to daughter cells requires a cytoplasmic pool of Nsp1. J Cell Biol 203:215-32 (2013). WB, ICC/IF . PubMed: 24165936
      • Ringel AE  et al. Yeast Tdh3 (glyceraldehyde 3-phosphate dehydrogenase) is a Sir2-interacting factor that regulates transcriptional silencing and rDNA recombination. PLoS Genet 9:e1003871 (2013). ICC/IF ; Saccharomyces cerevisiae . PubMed: 24146631
      • Beach A  et al. Mitochondrial membrane lipidome defines yeast longevity. Aging (Albany NY) 5:551-74 (2013). PubMed: 23924582
      • Motwani T  et al. Sir3 and epigenetic inheritance of silent chromatin in Saccharomyces cerevisiae. Mol Cell Biol 32:2784-93 (2012). ICC/IF ; Saccharomyces cerevisiae . PubMed: 22586263

      Customer reviews and Q&As

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      1-2 of 2 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-NSP1 antibody [32D6]

       Excellent
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Saccharomyces cerevisiae Cell (Yeast cells DDY1810)
      Permeabilization
      Yes - methanol/acetone
      Specification
      Yeast cells DDY1810
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50µg/mL · Temperature: RT°C
      Fixative
      Paraformaldehyde
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      Abcam user community

      Verified customer

      Submitted Sep 28 2010

      Question

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      Answer

      Thank you for your enquiry. I have contacted the source of the three antibodies and as such unfortunately I cannot give you an exact figure on the protein or IgG concentration of the antibodies. However, all three are derived from tissue culture supernatants and are therefore in the range of 0.5mgml-1 to 1.0mgml-1. Please do not hesitate to contact me should you require further assistance.

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