Nathan Soper

9 Questions 5 Answers 0 Followers

Questions related from Nathan Soper

EMSA: Why am I seeing bands below probe alone?

I am doing an EMSA experiment of protein binding to a fluorescent RNA probe. I am using a native polyacrylamide gel. The RNA is double stranded and I use ThermoFischer's protocol of annealing RNA...

26 February 2025 3,838 2 View

Is it possible for small molecule ligands to show significant heat release in control ITC run?

I am trying to optimize my control titration for ITC so that I can ensure my ligand to protein traces are reliable. My small molecule is stored in DMSO. I have been very careful to avoid buffer...

15 February 2024 9,178 3 View

Slight increase in heat transfer at the end of ITC trace?

Hello, I have recently switched from doing ligand into protein for ITC because my ligand is in DMSO and has low solubility at 10% DMSO; therefore, now I am doing protein into ligand -- the heat...

10 February 2024 5,071 1 View

Any tips on reverse ITC, protein into ligand?

Hello, I am having an ITC issue because I cant increase my ligand concentration due to solubility issues. I know there is past literature on reverse titrations being acceptable. I am more asking...

07 February 2024 5,611 2 View

How to rule out buffer mismatch in ITC Curve?

Hello, I am doing ITC and getting repeatable curves that look very promising. After reading some articles, it seems the biggest challenge to avoid is buffer mismatch. Is there a way to rule out...

19 January 2024 1,265 5 View

ITC Software not fitting correctly?

Hello, I have had moderate success with ITC but most of my issues have been with the Nano Program in fitting. I have a clear curve but the program is not fitting it correctly, does anyone have a...

23 December 2023 9,362 1 View

ITC Curve in the opposite direction?

Hello, I am doing a titration of ligand into protein and am getting good heat releases compared to my ligand into buffer. I am getting more heat release in later injections. Could cooperativity...

08 November 2023 799 4 View

Drop in Conductance when Using Size Exclusion?

Hello, I am using size exclusion to purify a protein. Before size exclusion a ran my sample through cation exchange. Where I typically see my protein come off during SEC, there was a drop in...

05 April 2022 231 6 View

Trouble Shooting New Cation Exchange Issue

Hello, I have been using cation exchange (HiTrap SP) for my purifications on ATKA. At that moment we only had two 1mL columns, one was HP and the other XL, and I was using those connected, and my...

01 January 1970 5,350 1 View