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
We are working on purifying a 60 kDa and have been having issues ridding the fractions of lower MW protein contaminants. We've previously used Ni-NTA and Ni Sepharose 6 fast flow resin for...
03 March 2019 8,285 5 View
Hello everybody, I want to purify a 32 kDa protein with a C-terminal His-tag. I use the Histrap Crude FF and I elute with 500 mM Imidazol (no linear gradient) and I get a quite distinct peak with...
03 March 2019 2,024 5 View
A recombinant his tagged protein (Size: 27 KDa Theoretical pI: 7.85) is eluted fine with 50 mM NaH2PO4 pH 8.0, 500 mM NaCl and 250 mM Imidazole. Precipitation occurs during dialysis with same...
03 March 2019 4,317 4 View
I work with the protein CWC2 which contains a zinc finger. My question is during the process of purification is zinc removed from the protein? If so, how should I go about re-adding it?
03 March 2019 7,950 3 View
Hi all, I am working with a protein that I have characterized quite well by SEC using superose6 resin. In the last 2 purifications, I have been unable to recover the protein in any fractions...
03 March 2019 4,811 6 View
Hi All, I have used MBP tag to help solubilize my target protein. It works really well. However, whenever I cleave off the His-MBP tag and try to clean up the sample by passing through another Ni...
03 March 2019 1,109 6 View
Can anybody explain what is the road map for solving Protein X-ray crystal structure? Thank you
03 March 2019 1,126 2 View
My model organism is Leishmania containing about 8000 total protein coding genes. I am performing LCMS based whole proteomics to do qualitative analysis under different conditions. What percentage...
28 February 2019 2,791 4 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...
21 February 2019 8,465 11 View
I want to do structural analysis of the protein after purification.
20 February 2019 3,879 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 1,828 14 View
I use the sonication method to lyse S. cerevisiae and quantify total protein extraction as a function of sonication time. Sometimes foam was formed, as the foam could mean protein...
13 February 2019 1,886 4 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 6,070 2 View
During animal cell culture, confluent cells are trypsinised, centrifuged, resuspended and finally subcultured. I read somewhere that contact inhibition is used to make the cultures synchronous....
04 February 2019 601 3 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 6,866 6 View
02 February 2019 7,324 15 View
02 February 2019 6,280 12 View
We are exploring the pathway of tumour formation in lung cancer. This lactic acid formation is done in mitochondria inner membrane in anaerobic conditions to form 2 molecules of ATP. # warburg...
26 January 2019 9,071 0 View
Do you know any FDA Approved drugs that I can purchase in order to induce H3K4-K9 methylation ?
19 January 2019 4,592 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 2,868 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 9,304 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 4,056 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 959 9 View
01 January 2019 4,105 8 View