Vikrant Kumar

6 Questions 7 Answers 0 Followers

Questions related from Vikrant Kumar

Intensity changes with probe - ligand complex vs probe only in FP assay?

I am using a chemical tagged with fluorophore for FP assay. The chemical is a known binder for the protein. After optimizing G-factor to 0.91 for improving the FP values, I observed in subsequent...

03 April 2023 3,942 5 View

Can racemic mixture behave different than components in a fluorophore biophysical assay? Can it be more active than pure fractions?

Biophysical assay involves cleaving a quenched fluoregenic substrate that results in increased fluorescence. Difference between racemic mixture and pure components came to be ten times different...

24 January 2023 9,699 1 View

In case of screening for inhibitors for an enzyme, is it important to pre-incubate the compounds with enzyme before adding the substrate? why?

I am used to pre-incubate the test compounds with enzyme for durations ranging from 15 min to 45 min before adding the substrate and initiating the reactions. However, in process of designing a...

13 December 2022 3,643 2 View

Is EC50 or EC80 concentraion of a ligand from a direct binding assay the better choice to establish a competitive assay as the next step?

The norm with most assays such as FP has been to use EC80, however with MST, the manual suggests different versions! Though it is simpler to establish a competition with EC50 concentration in...

03 September 2019 7,340 0 View

Is it correct to establish a competitive assay with two protein (hot and cold) vs a peptide (ligand)?

I want to understand the general principle while establishing a competitive assay. Should it be between a binding site vs two ligands (hot and cold peptides in case of peptide) competing for it or...

20 August 2019 1,547 2 View

Why would Kd value vary across FP, ITC and MST assay significantly?

I study protein-small molecule interaction. And it is a constant observation that I find. However referring to a literature I hardly come across such significant differences. What should be taken...

29 November 2018 2,005 5 View