Bruna Ukrainski

4 Questions 9 Answers 0 Followers

Questions related from Bruna Ukrainski

Why is my protein eluted with so many contaminants in Ni-NTA purification?

Hey, I need help! I'm trying to purify one proteins (82 kDa) using a Ni-NTA resin. The enzyme have a 6-histidine tag (bioinformatics models show that the tags are exposed correctly), and...

12 March 2025 915 5 View

HELP. Purification under denaturing conditions

Hello!! I have been working with a class of proteins that I cannot purify at all! They are all expressed in the soluble fraction, however, when we start purification, it comes out in the gradient...

01 January 1970 5,013 3 View

DTT may help remove non-specific interactions

Hello, everyone! I have seen many protocols for protein purification that use DTT or BME. I understand that it is to "break" disulfide bonds, but I would like to know what this implies in terms of...

01 January 1970 777 4 View

Why can't I transform my plasmid?

I'm working with the SSGCID plasmid (AVA0421), derived from pET 14b with amp resistance. I transformed this plasmid into DH5alpha and Rosetta (DE3) Plyss using amp 100 ug/mL, and it worked. Now...

01 January 1970 5,430 4 View