Nsp1p

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.\n\nApplications:\nSuitable for use in Immunofluorescence and Western Blot. Other applications not tested.\n\nRecommended Dilution:\nImmunofluorescence: For yeast cells, use diluted 1:1000-1:10,000.\nWestern 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.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

031414

Size

500ul

Applications

IF, WB

Hosts

Mouse

Form

Supplied as a liquid in 10mM sodium azide.

P Type

Mab

Purity

Sterile-filtered cell culture fluid from an Integra CL-350 biochamber

Isotype

IgG1

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).\n\n2. 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).

Additional Info

Recognizes yeast Nsp1p.