Recombinant RFP was synthesized with an amino terminal 6XHIS tag and was expressed in E. coli. A 3 step process was used to purify the protein: a partitioning 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 90% pure.
Form:
Liquid (sterile filtered)
Application Notes:
Application Notes: RFP Control is suitable as a control for polyclonal or monoclonal Anti-RFP in immunological assays. The molecular weight of RFP (calculated) is 27 kDa, but in a gel it is expected to run at approximately 28-30 kDa. Recombinant RFP has been tested in SDS-page, dot blot, and western blot
SDS-PAGE of purified RFP control shows a single band approximately 28-30 kDa (arrow). Approximately 1 µg of purified protein was loaded on to a 4-20% gradient gel for separation followed by staining with Coommassie blue. Molecular weight estimation was made by comparison to MW markers indicated at the right.
Western blot using RFP control protein. Lane 1: Super Signal Molecular Weight. Lane 2: 50 ng of RFP (p/n orb345960). Primary Antibody: Rabbit Anti-RFP (p/n orb345391) at 1:1000 overnight at 2-8C. Secondary Antibody: Goat Anti-Rabbit IgG HRP (p/n orb347654) 1:40000 for 30 mins at RT. Expect ~27kDa.
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