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Immobilisation level
- OldForum
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12 years 8 months ago #1
by OldForum
Immobilisation level was created by OldForum
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
Thank you
Kati
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- OldForum
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12 years 8 months ago - 9 years 8 months ago #2
by OldForum
Replied by OldForum on topic Immobilisation level
Suggestion:
www.sprpages.nl/immobilization.html
Last edit: 9 years 8 months ago by .
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- OldForum
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12 years 8 months ago #3
by OldForum
Replied by OldForum on topic Immobilisation level
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
all best,
eya
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