Nsp1p

Référence 031414-500ul

Conditionnement : 500ul

Marque : US Biological

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031414 Nsp1p

Clone Type
Monoclonal
Host
mouse
Swiss Prot
G2WGX7
Isotype
IgG1
Grade
Supernatant
Applications
IC IF IHC IP WB
Crossreactivity
Ye
Shipping Temp
Blue Ice
Storage Temp
-20°C

This antibody recognizes Nsp1p, a nuclear pore complex protein, and is a useful marker of yeast nuclear pores. 031414 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 encoding ~66% of the C-terminal portion of Nsp1p. To confirm that 031414 possessed high affinity for Nsp1p, strain RS453, which expresses a shortened isoform of Nsp1p, was compared to the wildtype strain BJ5465. NSP1 in the RS453 strain contains an internal deletion that removes the coding sequence for 6 FXFG repeats, which are not essential for protein function, and encodes a protein that has been observed to migrate at approximately 85kD on SDS-PAGE gels. The predicted size of wildtype Nsp1p is 86.5kD, but Nsp1p has been observed to migrate on SDS gels at ~100kD. In our SDS-PAGE gels, the apparent molecular mass of wild type Nsp1p was 108kD, whereas the isoform of Nsp1p expressed in the RS453 strain migrated at 91kD. The detection of two protein bands of apparent sizes 108kD and 91kD in the BJ5465 and RS453 strains, respectively, demonstrated that 031414 recognized the pore complex protein Nsp1p.

Applications:
Suitable for use in Immunofluorescence/Immunocytochemistry, Immunohistochemistry, Immunoprecipitation and Western Blot. Other applications not tested.

Recommended Dilution:
Immunofluorescence: For yeast cells, use diluted 1:1000-1:10,000.
Western Blot: For yeast protein samples, use diluted 1:10,000 (cell lysates) to 1:50,000 (nuclear fractions), followed by chemiluminescent detection (ECL). For other (non-ECL) Western detection methods, try diluted 1:1000-1:5000.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Applications
Product Type: Mab|Isotype: IgG1|Clone No: 32D6|Host: mouse|Concentration: Not Determined |Form: Supplied as a liquid, 5mM sodium azide.|Purity: Supernatant|Immunogen: Yeast nuclear preparations|Specificity: Recognizes Sacchoromyces cerevisiae Nsp1p.||Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Immunogen
Yeast nuclear preparations
Form
Supplied as a liquid, 5mM sodium azide.
Purity
Supernatant
Specificity
Recognizes Sacchoromyces cerevisiae Nsp1p.
References
1. Tolerico, L. H., A. L. Benko, J. P. Aris, D. R. Stanford, N. C. Martin, and A. K. Hopper. Saccharomyces cerevisiae Mod5p-II contains sequences antagonistic for nuclear and cytosolic locations. Genetics 151:57-75 (1999).||2. Fahrenkrog, B., J. P. Aris, E. C. Hurt, N. Pante, and U. Aebi. Comparative spatial localization of protein-A-tagged and authentic yeast nuclear pore complex proteins by immunogold electron microscopy. J. Struct. Biol. 129:295-305 (2000).