Recombinant Red Fluorescent Protein (RFP) Control

Recombinant RFP was synthesized with an amino terimal 6XHIS tag and was expressed in E.coli. A 3 step process was used to purify the protein: a partioning separation using t-butanol, followed by hydrophobic interaction chromatography (HIC), and lastly IMAC nickel chromatography. Analysis by SDS-PAGE resulted in a pattern consistent with purified RFP and was estimated to be greater than 95% pure. Recombinant RFP (wild type, sequence corresponding to RFP, Discosoma spp.) is intended for use as a control when using polyclonal or monoclonal anti-RFP in immunological assays, in fluorometry, and in fluorescence activated cell sorting (FACS). Polyclonal anti-RFP can detect RFP by ELISA and western blot. Biotin conjugated polyclonal anti-RFP used in a sandwich ELISA is well suited to titrate RFP in solution when used in combination with unconjugated anti-RFP antibody. This product can be used as a control in western blots.