Hi,
I have been tryig to get curvature sensogram using a proteinA chip for single cycle kinetic but instead of it the sensogram raising linearly and reaching quickly to steady state just after the injection of the each dilutions. I use the parameters below,
Dissociation time: 900s
Analyte flow rate: 30ul/min
Running buffer: HBS-EP+_1M NaCl
Concentrationrange: 3nm to 0.037nm
i have already tried very different range of analyte concentrations.
To clearfy my question please see the photo of the sensogram.
Dear Nil,
the best thing to do is to further reduce the ligand density of your surface to get lower Rmax. The other question however is whether you really need "more" curvature. Some interactions are fast and tight and in those cases you will not see much curvature under most experimental conditions. But depending on what you want to know you may not absolutely need it. If you need it, then you have to reduce the surface capacity. And you can try to run the experiment at 37°C to get a faster off-rate. However, looking at your sensorgram i think the off-rate is fast enough to get a reliable fit.
Cheers Markus
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