My data points show a both static and dynamic quenching mechanism, but I can't fit it to a polynomial grade 2 equation to extrapolate the values to the Stern-Volmer derived equation F0/F = 1 + (KD+Ks)[Q] + KDKs[Q]2. I am using OriginPro8. I appreciate your help, thank you in advance.
The name of the Great Oracle: Google
and the keywords: OriginLab nonlinear fitting
in Mathematica, use monte carlo.
https://arxiv.org/ftp/arxiv/papers/1105/1105.6254.pdf
Are you sure you have both types of quenching in your system under investigation?
Do you have any possibilities to do lifetime measurements?
Fistly, you will prove the dynamic type of quenching. Secondly, you can try to extract the K(dynamic) from that and use it to fit the data more precisley!
Another thing is that you have probably two different fractions of the fluorophore accessible to the quencher.
Nikita,
thank you for your answer. I'm assuming both types of quenching based on the plot F0/F vs [Q] since it shows an upward curvature as described by Lakowicz. How can I test for the accessibility of the quencher ? Does it depend on the number of binding sites and stoichiometry for example ?
Thank you.
Hi Sudarson,
here's the scatter plot of my data. Thank you for your help.
Dear Satish,
I was able to fit the data to Stern-Volmer equation and extract Ksv values.
Please check my publication (dx.doi.org/10.1016/j.biochi.2017.02.005) to find the software and equations used.
Josué
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