A forum to address questions regarding methods of protein purification. | Contact experts in Protein Purification to get answers
1,187 views 1,789 posts
Questions related to Protein Purification
Do you know any FDA Approved drugs that I can purchase in order to induce H3K4-K9 methylation ?
19 January 2019 4,531 3 View
I have to check the methylation activity of my protein so i referred your paper JMJ14 is an H3K4 demethylase regulating flowering time in arabidopsis (Cell Research). so can anyone send me the...
01 January 2019 2,818 4 View
Protein was expressed in SHuffle cells in presence of 0.5%glycerol in the LB media(230ml). pellet was dissolved in 20ml lysis buffer(50mM Tris pH 8.0, 150mM Nacl, 10% glycerol). i couldnt see...
01 January 2019 9,250 7 View
I have been attempting to purify a His-tagged bacterial chaperonin protein. I have had no issue over-expressing the protein; however, it appears my protein is not binding well enough to my column...
01 January 2019 4,017 8 View
hey i am using 2%OG and 1%DDM what concentration do i need to use with DM? is there a formula to calculate? Why usualy using 1%DDM and 2% OG (what % is higher then the CMC?) thanks
01 January 2019 902 9 View
01 January 2019 4,014 8 View
I have expressed a human glycoprotein in plant ER through a signal sequence and it also contains ER retention signal at the C-terminus. Thus, if this protein contains an N-glycan moiety, it should...
20 December 2018 4,442 3 View
I am studying effect of stress on protein-protein interaction. For that control cells are stored at -80C without any stress. However, Before performing protein purification, Can cells be stored...
13 December 2018 4,255 1 View
12 December 2018 9,445 3 View
My project is purification protein and when I change solution by dialysis, my protein is precipitated at the time pH of solution is pI. So how to protein can dissolve again? Thanks you!
12 December 2018 6,351 4 View
Hi all, I am working on a tricky protein. I surpassed some initial aggregation behaviour by adding 0.3% CHAPS and 1mM TCEP, buffer in which is happy until the removal of the SUMO tag, then it...
12 December 2018 2,859 3 View
I purified a protein in BL21 pLys, S (E. coli strains). After purification, the protein was degraded. How can I solve this problem?
12 December 2018 5,942 0 View
10 December 2018 9,585 3 View
I am going to purify several proteins by using GFP-nanobodies linked to a sepharose column since my proteins have a GFP taq. Since I have not experience with this protocol, I would like to solve...
21 November 2018 1,735 1 View
11 November 2018 6,210 1 View
I produced recombinant βA domain of β-Integrin protein from shrimp using recombinant plasmid pGEX4T1- βA. The plasmid was transformed into E. col BL21 Star. The cells were induced by 0.5 mM IPTG....
11 November 2018 3,974 8 View
Is it possible to make an Ni-NTA affinity column with a syringe? if so how? My column is blocked and no solution is eluted from it. So i can't purify my His-tagged protein using this column. I...
11 November 2018 7,784 4 View
Protein purification percent yield percent recovery
10 October 2018 9,492 3 View
Hi all, a question for the protein purification masters here. After protein elution from a Ni column, some people simply dilute the sample down to decrease the imidazole concentration. I have...
10 October 2018 2,532 10 View
Hi all, I have this his-tagged protein which I wanna pull down and want to perform a Chip-seq. While Ni-NTA is used for a lot his-tagged protein purification, I didn't find any citations using...
10 October 2018 8,162 4 View
I have get insolube protein at 1mMIPTG after 4 hours induction time. How can I find solube protein for purification by his tag nikal colum.
10 October 2018 8,451 6 View
I am trying to purify a membrane protein and a transcription factor. We are planning to use Step tag for purification. Can someone shed light on this method? Is it better than traditional GST and...
10 October 2018 5,625 0 View
Dear friends, We have a bunch of small molecules (around 20) and purified protein as potential target. I am wondering what is the fastest and cheapest experiments to find if there is any...
10 October 2018 3,725 28 View
Hi guys, I am wondering if anyone has using TB instead of LB for the pre culture of isotope labeled protein (C13 and N15) and what the yield is like compare to LB transfer. More precisely, my lab...
10 October 2018 7,234 4 View