Analyte buffer - Forum

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This forum is intended for questions about kinetics, Surface Plasmon Resonance and the instruments related to these techniques.
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TOPIC: Analyte buffer

Analyte buffer 07 Sep 2017 16:45 #1

Just want to say this website is a fantastic resource, thank you !

I wanted to know the best way to deal with analyte buffer and running buffer discrepancies to get the best I know they should be matched but they can't be buffer exchanged
I am running purified antibody samples from various subjects for kinetic analysis
I run each at 5 concentrations in a serial dilution
Each of my analyte/antibody is at different stock concentration in PBS
I dilute them in my running buffer
Low conc stock analyte means almost 75% of my injection will be PBS for that sample at the highest conc
Will my reference flow cell subtraction take care of this bulk effect?
Or should I run a blank of not running buffer but of the concentration of pbs in running buffer for my highest conc to double ref for each sample?

Thanks! Hope this makes sense!

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Analyte buffer 08 Sep 2017 08:27 #2

Thank you for your kind words.

If your analyte is in PBS, why not using PBS as running buffer? I cannot think of a reason why to stick to the HBS buffer.

In general small buffer mismatches will be compensated by reference subtraction and double referencing should remove most of the differences.

Kind regards

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