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. Author manuscript; available in PMC: 2011 Jan 21.
Published in final edited form as: Methods Mol Biol. 2010;627:75–90. doi: 10.1007/978-1-60761-670-2_4

Figure 1. A typical single flow cell sensorgram generated during the loading of anti-GST antibody.

Figure 1

The flow cell surface was activated for direct amine coupling of the anti-GST antibody (αGST-Ab) by injecting the coupling solution (40 μl NHS/EDC at 5 μl/min flow rate with EXTRACLEAN command). Anti-GST antibody was then diluted to 30 μg/ml in immobilization buffer and 45 μl were injected at 5 μl/min flow rate, omitting the EXTRACLEAN command. To deactivate esters on the sensor chip surface, ethanolamine was injected (35 μl at 5 μl/min flow rate with EXTRACLEAN command). To test the binding surface, supplied recombinant GST protein was diluted to 5 μg/ml and 100 μl were injected over the sensor chip at 20 μl/min flow rate. To remove GST from the binding surface, 40 μl of glycine-HCl pH 2.2 was injected at 20 μl/min flow rate.