Does low pH affect MP readings?
I am taking MP readings of my protein of interest at 100nM in varying buffer pHs (9.0 to 6.0). The readings are successful until about a buffer pH of 6.0, in which the number of events drastically decrease (from about 1000-1500 at pH 6.5, to about 150-200 at pH 6.0). Are there known influences of low pH buffers on MP readings?
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Hi Keith,
As long as the low pH does not degrade the target molecules, then the mass will be the same. You are also changing the net charge of the protein so hopefully it will still bind to the negatively charged glass… otherwise you will need to coat slide with poly lysine.
However you should use the appropriate buffer for calibration as well as for the measurement.
Folded/unfolded proteins will have the same mass and our technique is insensitive to protein confirmation/orientation.
Hope that helps!