Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Western Blotting (WB)
Aufreinigung
conc. tissue culture supernatant
Sterilität
Sterile filtered
Klon
32D6
Isotyp
IgG3
Applikationshinweise
For western blots of yeast protein samples, use MCA-32D6 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 MCA-32D6 diluted 1/1,000 to 1/5,000. For immunofluorescence on yeast cells, use MCA-32D6 diluted 1/1,000 to 1/10,000. Antibody preparation contains 10mM sodium azide preservative .
Beschränkungen
Nur für Forschungszwecke einsetzbar
Format
Liquid
Konzentration
1 mg/mL
Buffer
It is supplied as a sterile-filtered cell culture fluid from an Integra CL-350 biochamber plus sodium azide. The antibody class is IgG1 and the antibody concentration is about 1mg/mL.
Konservierungsmittel
Sodium azide
Vorsichtsmaßnahmen
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung
Avoid repeated freezing and thawing.
Lagerung
4 °C/-20 °C
Informationen zur Lagerung
Store at 4°C short term or -20°C long term.
Target
NSP1
(Nsp1p (NSP1))
Andere Bezeichnung
Nsp1p
Hintergrund
MCA-32D6 recognizes Nsp1p, a nuclear pore complex protein, and this antibody is a useful marker of yeast nuclear pores. The monoclonal antibody MCA-32D6 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 approx. 66% of the C-terminal portion of Nsp1p. To confirm that MCA-32D6 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 85 kDal on SDS-PAGE gels. The predicted size of wildtype Nsp1p is 86.5 kDal, but Nsp1p has been observed to migrate on SDS gels at approx. 100 kDal. In our SDS-PAGE gels, the apparent molecular mass of wild type Nsp1p was 108 kDal, whereas the isoform of Nsp1p expressed in the RS453 strain migrated at 91 kDal. The detection of two protein bands of apparent sizes 108 kDal and 91 kDal in the BJ5465 and RS453 strains, respectively, demonstrated that MCA-32D6 recognized the pore complex protein Nsp1p.