Forum :: Immobilisation level


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TOPIC: Immobilisation level

Immobilisation level 24 Feb 2012 01:00 #1

Hi, I wonder what is the proper immobilization level for my experiments in Biacore. I was using about 1000 RU and it was ok for kinetics measurements. However, a different protein was immobilized at about 1000 RU also and it did not work at all, I mean protein was immobilizzed but no binding was observed... Any suggestion??
Thank you
Kati

Immobilisation level 24 Feb 2012 01:00 #2

Last Edit: 13 Mar 2015 21:27 by MQAdmin03.

Immobilisation level 24 Feb 2012 01:00 #3

You should always take the molecular mases of your molecules into the account. Use formula Rmax= MWanalyte/MWligand*Rimm*S, where S is stoichiometric ratio and Rimm is immobilization level. Adjust Rimm to obtain Rmax (maximum binding of analyte) values between 50-100 RU (for kinetic analysis).
all best,
eya