Application
Western blots of whole yeast protein extracts with a collection of our antibodies. The blot for is in the indicated lane, and the number indicates the SDS-PAGE molecular weight in kDa.| Product Name | Nsp1p antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mouse monoclonal [32D6 ] to Nsp1p - Nuclear Envelope and Nuclear Pore Marker |
| Immunogen | This was raised against a yeast nucleolar prep, and screened by immunocytochemistry and western blotting. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Clone Number | 32D6 |
| Host Species | Mouse |
| Tested Applications | ICC/IFWB |
| IF: Use at a dilution of 1/1000 - 1/1000 (yeast cells). WB: Use at a dilution of 1/10000 (cell lysates) - 1/50000 (nuclear fractions). For other (non-ECL) detection methods, use at a dilution of 1/1000 - 1/5000. Detects a band of approximately 108 kDa and 91 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
|
| Concentration | 1mg/ml |
| Target | Nsp1p |
|---|---|
| Tissue 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 Nsp1p, strain RS453, which expresses a shortened isoform of Nsp1p, 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 Nsp1p is 86.5 kDa, but Nsp1p has been observed to migrate on SDS gels at ~100 kDa. In our gels, the apparent molecular mass of wildtype Nsp1p was 108 kDa, whereas the short isoform of Nsp1p 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 Nsp1p. |
| Cellular Localization | Nuclear (pore complex and envelope) |
Application
Western blots of whole yeast protein extracts with a collection of our antibodies. The blot for is in the indicated lane, and the number indicates the SDS-PAGE molecular weight in kDa.| Application Notes | IF: Use at a dilution of 1/1000 - 1/1000 (yeast cells). WB: Use at a dilution of 1/10000 (cell lysates) - 1/50000 (nuclear fractions). For other (non-ECL) detection methods, use at a dilution of 1/1000 - 1/5000. Detects a band of approximately 108 kDa and 91 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | Preservative: 10mM Sodium Azide; Constituents: Tissue Culture Supernatant |
