Fol. Biol. 2005, 51, 50-51
Specification of the Monoclonal Antibody PK1 Reactivity in Chinese Hamster Ovary Cells
eIF3a/Rpg1p/Tif32p is the evolutionarily conserved subunit of yeast eukaryotic translation initiation factor 3 (eIF3) that promotes multiple steps of the initiation pathway resulting in formation of the 48S pre-initiation complex at the AUG start codon of mRNA. The monoclonal antibody PK1 (IgG1 kappa) (Jiřincová et al., 1998) against the recombinant protein Rpg1 (Kovarik et al., 1998) has already proved itself as a useful tool for analysing the eIF3 complex in S. cerevisiae (Valášek et al., 1998). It binds either to native or denatured epitopes of Rpg1p and it can be used to efficiently isolate the eIF3 complex by immunoprecipitation (Valášek et al., 1999). This antibody was also used to show by immunofluorescence that yeast eIF3a is a microtubule-interacting protein (Hašek et al., 2000). In order to test whether this characteristic of yeast eIF3a is also conserved in higher eukaryotes, we analysed the immunoreactivity of the anti-Rpg1p monoclonal antibody (PK1) in Chinese hamster CHO, human HeLa and mouse 3T3 cells.
Funding
This work was financed by grant GACR 204/02/1424 to J. H. and by Institutional Research Concept No. AV0Z5020903.
References
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