A forum to address questions regarding methods of protein purification. | Contact experts in Protein Purification to get answers
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Questions related to Protein Purification
I want to do structural analysis of the protein after purification.
20 February 2019 3,757 6 View
I am currently trying to run dialysis on a protein purified using the ProBond Ni-Ta purification columns against a buffer of 10mM TRIS (pH 8.0) and 0.1% Triton X100 in sterile millipore water. My...
15 February 2019 1,729 14 View
Hi fellow researchers, for a project that i'm currently performing an protein purification step is required. For this we already have a recombinant protein fused to the Fc part of an...
04 February 2019 6,013 2 View
Dear All, Can we incubate Supernatant and Ni charged beads for binding at room temperature for overnight? Currently, we do not have a fixed temp room in our building. What about keeping Ni...
02 February 2019 6,779 6 View
02 February 2019 7,249 15 View
I am looking for the protein expression of Methanosarcina mazeii that can get expressed in the presence of acetate. I am not much aware of steps. I was planning to do cell-lysis and then SDS-PAGE...
02 February 2019 6,164 12 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 2,809 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 9,239 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 4,007 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 889 9 View
01 January 2019 3,997 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 4,428 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 4,230 1 View
12 December 2018 9,428 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 6,339 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 2,847 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 5,935 0 View
10 December 2018 9,563 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 1,717 1 View
11 November 2018 6,192 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 3,967 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 7,778 4 View
Protein purification percent yield percent recovery
10 October 2018 9,485 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 2,517 10 View