Forum :: Analyte buffer


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

TOPIC: Analyte buffer

Analyte buffer 07 Sep 2017 16:45 #1

Hi
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!

Analyte buffer 08 Sep 2017 08:27 #2

HI,
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
Arnoud
Moderators: Arnoud