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. Author manuscript; available in PMC: 2014 Nov 19.
Published in final edited form as: Anal Chem. 2013 Oct 28;85(22):10.1021/ac402871k. doi: 10.1021/ac402871k

Figure 3.

Figure 3

Effect of calcium on the efficiency of protein ligation. A GFP-STEPL fusion protein was expressed and washed on an affinity column. The column was then treated with various concentrations of Ca2+, in the presence of triglycine, at 37°C for 4hrs. These experiments provided a measure of the total amount of ligated and unligated product ([GFP]Total) released from the affinity column for each Ca2+ concentration. The amount of unligated product ([GFP]unligated) was determined by performing analogous experiments in the absence of triglycine. The percent protein conjugated ([GFP]ligated) was then calculated calculated as ([GFP]Total − [GFP]unligated)/([GFP]Total)*100.