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. Author manuscript; available in PMC: 2009 Mar 5.
Published in final edited form as: Biochemistry. 2008 Aug 9;47(35):9098–9106. doi: 10.1021/bi800938g

Figure 7.

Figure 7

Purification of recombinant RS1 from Sf21 insect cell medium. A. Cell medium from Sf21 insect cells expressing RS1 was applied to a DEAE-Sephacryl column and bound protein was eluted with 20 mM TrisCl buffer, pH 7.5 containing increasing concentrations of NaCl. B. The fraction that eluted from the anion exchange column with 300 mM NaCl was applied to a galactose-agarose column. After removing the unbound protein, bound RS1 was eluted 6 times with 1 M IPTG with the majority of RS1 present in elutions 1–3. Samples from the anion and affinity columns were analyzed on SDS gel stained with Coomassie Blue to detect total protein and Western blots labelled with the RS1 3R10 antibody to detect RS1. C. Purified RS1 migrates as a 186 kDa octameric complex as revealed on SDS gels stained with Coomassie Blue.