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 am working with a protein-12kda; my vector contains 10X His tag My protocol for purification- NOTE- this is from 1 ml sonicated supernatant, used 100ul NiNTA beads. After I collect Flow...
24 May 2019 9,206 8 View
Can a human recombinant glycoprotein produced in mammalian cell culture lose part of its sialic acid residues due to changing conditions in the culture or the purification of the protein? Any...
18 May 2019 10,037 3 View
I express my protein in pTYB12 vector as fusion with intein and then try to purify on chitin beads. Typically, I load overnight at low flowrates (something like 0.3 ml/min). The loading buffer is...
09 May 2019 8,639 5 View
05 May 2019 8,469 9 View
I am looking for suggestions or comments from protein purification community, especially people who work with Con-A column chromatography. I am used Con-A Sepharose and Con-A agarose beads...
05 May 2019 9,939 6 View
I am trying to extract the protein content of extracellular bone matrix. After milling the frozen bone in nitrogen liquid, I have used RIPA buffer and HALT to extract the protein content, but...
05 May 2019 6,133 4 View
I am curious about nickel-bead cleaning. After native protein purification, I washed the nickel bead with 2 column volume of 2M Imidazole to remove the non-specific binding protein. But after...
05 May 2019 4,624 8 View
05 May 2019 5,264 3 View
05 May 2019 2,717 7 View
I am doing some his-tagged protein purification and I would like to see if the chromatography works by running a standard. Is there a cheap histidine tagged protein people tend to use? Also the...
08 April 2019 9,404 4 View
Hi, I'm looking to further purify my protein by using cation exchange with Nuvia S resin. However, my lab doesn't have an IEX system. Is it possible to still carry out a cation exchange...
04 April 2019 7,020 5 View
I have some questions regarding the AKTA pure system, mainly about the chromatogram.
04 April 2019 2,792 4 View
04 April 2019 2,596 5 View
Hi, I am working with a protein that is typically secreted from the parasitic cells that I study. I am trying to express and purify it from HEK cells. Despite of using the mammalian signal...
02 April 2019 7,272 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...
18 March 2019 7,829 2 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...
09 March 2019 5,495 5 View
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 8,229 5 View
03 March 2019 1,972 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 4,269 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 7,893 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 4,745 6 View
03 March 2019 1,008 6 View
Can anybody explain what is the road map for solving Protein X-ray crystal structure? Thank you
03 March 2019 1,077 2 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 8,384 11 View