Molecular and Biochemical Characterization of Recombinant Guinea Pig Tumor Necrosis Factor-Alpha

Figure 1

(a) Coomassie blue-stained SDS-PAGE analysis of eukaryotic protein elutions from cells transfected with 18 μg of pCEP-Pu plasmid DNA without any gene (vector control) or pCEP-Pu plasmid DNA (18 μg) containing TNF-α gene. The arrow indicates a ~19 kDa band which was recognized as rgpTNF-α. Absence of band in the vector control is also shown. MM: molecular marker; V: vector control elutions, and T: TNF-alpha elutions. (b) Coomassie blue-stained SDS-PAGE analysis of prokaryotic expressed rgp-TNF-α. rgpTNF-α was expressed using prokaryotic expression system (pQE-15) and analyzed on SDS-PAGE gel. MM: molecular marker. (c) Identification of eukaryotic expressed rgpTNF-α by polyclonal antiserum (1 : 2000 dilution) from immunized rabbits. Approximately 200 nanograms of rgpTNF-α was run on 10–20% tricine gel and analyzed by western blot analysis for checking its specificity. The ladder was stained with Coomassie Brilliant Blue.

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