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Questions related from Sam Mahdi
Hello everyone, I know about the superdex column turning yellow overtime, however the issue I'm having is it turned yellowish-brown AFTER 0.5M Sodium Hydroxide wash (2 CVs). This is a Superdex...
31 August 2020 5,094 0 View
Hello everyone, I've written a basic program to calculate the pI and MW of my protein (didn't like the format of ProtParam, so decided to make my own. However, the pI and MW calculated from my...
02 July 2020 9,655 2 View
I have a protein construct in a pETDuet vector that I mutated in a TEV cut site (ENLYFQS) between the His-Tag-BamHI cutsite, and my protein of interest. Sequencing confirmed proper sequence (e.g....
02 January 2020 3,694 2 View
In short, ideally the cobalt resin can be reused multiple times without any problems; however, reducing agents can damage the column (chelate the cobalt). Both personal experience, and from...
29 April 2019 3,395 0 View
Hello everyone, How does one determine the concentration of a sample with multiple proteins? I.E. If you have a complex that is formed from 3 different proteins, how do you determine it's...
31 January 2019 4,038 10 View
Hello everyone, I accidentally got my buffers mixed up, and added a buffer containing 10mM DTT to my cobalt column during my purification. Luckily the immediate color change indicated this mix...
29 December 2018 7,092 0 View
Hello everyone, I know one can calculate the theoretical pI based on the primary structure (i.e. sequence), but I also know this is slightly inaccurate due to how the protein folds (as indicated...
20 April 2018 1,837 4 View
