Less point in dissiciation phase - 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: Less point in dissiciation phase

Less point in dissiciation phase 23 Sep 2015 10:58 #1

  • Monica#1
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Hello Everyone,
I have been studying binding kinetics with BiacoreX100. The problem is that the dissociation phase drops sharply in the curves and there are not enough points in dissociation point so the fitting is really bad. I have tried with increased dissociation time from 200 sec to 600 sec but in vain. Pls see the attached figure.
Details: Chip -NTA
Buffers: HEPES buffer with Surfectant.
Flow rate for Ni-10uL/min
Flow rate for ligand: 5uL/min
I really appreciate any help/suggestions.
Thanks
Monica
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Last edit: by Monica#1. Reason: Addition

Less point in dissiciation phase 24 Sep 2015 10:12 #2

Hi Monica,

Is it possible to set the sampling rate to high with the X100? Then you have more points to analyse. Longer dissociation will not help you since the information of the dissociation is in the first few seconds because it is so fast.

An alternative is to do equlibrium analysis since you have equilibrium in all curves. This will give you de KD but not the kinetics.

Third option is to use another software package.
I can recommend Scrubber ( www.sprpages.nl/software-weblinks ). It is not free, but good value for money.

Kind regards

Arnoud

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Last edit: by Arnoud.

Less point in dissiciation phase 27 Sep 2015 17:35 #3

  • Monica#1
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Hi Arnoud,
Thanks for the reply. Could you pls elaborate on seting sampling rates to high. I am Not sure I get it.
Thanks again
Monica

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Less point in dissiciation phase 27 Sep 2015 21:41 #4

Hi Monica,

I know form the Biacore 2000 en T200 that it is possible to set the data sampling rate to high (10Hz) which means that you have more data points to analyse.

I am not sure that it is possible with a X100. I did not find anay reference to it.

Arnoud

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Last edit: by Arnoud.

Less point in dissiciation phase 30 Sep 2015 10:18 #5

  • Monica#1
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Hi Arnoud,
I don't think it is available in BiacoreX100. Thanks for your help.
Regards
Monica

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