Sp1p antibody

Catalog No. ABIN1842293

Product Details

Target: Sp1p

Reactivity: Saccharomyces cerevisiae

Host: Mouse

Clonality: Monoclonal

Application: Western Blotting (WB)

Specificity: Yeast nuclear pore complex protein, useful nuclear pore complex and nuclear envelope marker.

Characteristics: 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 A00109.01 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 approximately 85,000 Da on SDS gels. The predicted size of wildtype Nsp1p is 86,500 Da, but Nsp1p has been observed to migrate on SDS gels at ~100,000 Da. In our gels, the apparent molecular mass of wildtype Nsp1p was 108,000 Da, whereas the short isoform of Nsp1p in RS453 migrated at 91,000 Da. The detection of two protein bands of apparent sizes 108,000 Da and 91,000 Da in BJ5465 and RS453, respectively, demonstrated that A00109.01 recognized the pore complex protein Nsp1p.

Purification: Immunoaffinity purification

Immunogen: YA32D6 recognizes Nsp1p, a nuclear pore complex protein, and this antibody is a useful marker of yeast nuclear pores.

Isotype: IgG

Application Details

Application Notes: Western blots of yeast protein samples: 1:5,000 (cell lysates) to 1:25,000 (nuclear fractions), followed by chemiluminescent detection (ECL).
Other (non-ECL) Western detection methods: 1:500 to 1:2, 500
Immunofluorescence on yeast cells: 1:500 to 1:5,000

Restrictions: For Research Use only

Handling

Format: Frozen

Concentration: 1 mg/mL

Buffer: PBS, pH 7.4 with 0.05 % sodium azide. frozen liquid

Preservative: Sodium azide

Precaution of Use: WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.

Handling Advice: Briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container's cap because small volumes of antibody will occasionally become entrapped in the seal of the product vial during shipment and storage.
Avoid repeated freezing and thawing.

Storage: -20 °C

Storage Comment: Stable for 1 year at -20 °C from date of shipment. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquot to avoid repeated freezing and thawing. Aliquot will be stable at 4 °C for 3 months.

Target Details

Target: Sp1p