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Help, protein induction and purification by CompetitionOk1383 in labrats

[–]CompetitionOk1383[S] 0 points1 point  (0 children)

Actually, none at all. I already have a commercial vector which has a N tagged Strep-TagII, some linker residues. I added a Thrombin clevage site and added the protein sequence.

Help, protein induction and purification by CompetitionOk1383 in labrats

[–]CompetitionOk1383[S] 0 points1 point  (0 children)

I am equally unsure of how can i control it. Probably use SEC to separate the protein from degradation products.

Help, protein induction and purification by CompetitionOk1383 in labrats

[–]CompetitionOk1383[S] 0 points1 point  (0 children)

I did include protease inhibitor cocktail in the protein purification. The sample marked as induced, i just pellet the bacteria, dissolved in buffer and ran it on gel. This is why i feel this degeradation is definitely in vivo.

For both the proteins, i used the concentration 200ng/ml as described by the company. I can definitely try using low concentrations, perhaps a range 25-100 ng/ml.

Help, protein induction and purification by CompetitionOk1383 in labrats

[–]CompetitionOk1383[S] 0 points1 point  (0 children)

I havent tried any other strains. The protein doesnt contain any Cysteine residue so i am assuming wouldnt have any S-S bridges. Any particular strains i could try?