I am titration protein in the RNA sample. Titration was performed in binding buffer, RNA in the cell was 2.5uM and protein in the syringe was 96uM, with 19 injections (2uL, 4s. I have attached the file below.
If anyone could help, that would be easier for me to proceed for optimization.
Thank you in advance.
Dear Ajit Kumar
One reason for such signal may be aggregation/precipitation of the protein-RNA complex. If this process is time-dependent, shortening the titration time may help (probably, you can do it by shortening the time between injections by approx. 20%). Additionally, you can lower the concentrations (also by approx. 20%). Slowing down the mixing is another thing that comes to my mind (too fast mixing tends to denature proteins).
The second approach would be doing the reverse titration, i.e., RNA to protein.
The next reason for such signal may be a too big air bubble at the top of the syringe. Injection of the air instead of the sample solution would destroy the result.
I hope it helps.
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