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Questions related from Michele Costantino
I've recently purified TEV with a his-tag on a Ni-NTA column using our AKTA Pure FPLC. It usually works great. This time I had two peaks of lower absorbance over many fractions in the elution...
28 August 2023 7,374 0 View
I have been modifying a protein through a radical reaction, which conjugates to tyrosine. As such, measuring protein concentration by absorbance at 280 nm is not working anymore. So I was told,...
15 June 2022 5,079 4 View
I am working with a protocol that requires cell pellet resuspension in 6 M guanidinium. I did not make this protocol, so I don't know how they came to this optimization. I keep getting very...
09 December 2021 6,844 2 View
I'm training an undergrad and they didn't clamp the dialysis bag correctly. All the protein leaked out in the dialysis buffer. I now have my 15 mL sample floating around in 2 L of buffer. The...
15 April 2021 9,631 3 View
I have been expressing a protein with a tag which is then removed with a protease and repurified. Both purification steps use a Ni-NTA column. I run the samples on SDS-PAGE (15%, self-poured)....
28 December 2020 7,400 1 View
