A forum to address questions regarding methods of protein purification. | Contact experts in Protein Purification to get answers
1,973 views 1,849 posts
Questions related to Protein Purification
I'm extracting protein from highly hazardous bacterial samples. For the safe of safety, people commonly boil this bacteria prior to full lysis when doing protein extraction. Because the bacteria...
03 March 2020 966 8 View
Hi, I routinely use AKTA connected to Fraction Collector-920 for collecting proteins while doing protein purification. However, recently I have started seeing some errors on our Fraction...
28 February 2020 9,242 4 View
I am studying a single-chain variable fragment (scFv) linked to a luciferase (Nanoluc) at the C-terminus (N-scFv-linker-Nluc-C). The naked scFv (not linked to Nluc) can be easily purified using...
23 February 2020 9,206 2 View
In order to keep my Tris 20 mM GSH pH8 elution buffer active for as long as possible I would like to know if I could prepare a large quantity and freeze it? (how much time can I freeze/thaw it?)...
20 February 2020 8,527 3 View
I am trying to purify a cysteine protease to use in an activity-based assay. The gene has been sequenced. It is in a pET based vector and has a His6 and GFP tag. I express in BL21 DE3 and the...
02 February 2020 678 8 View
02 February 2020 6,284 2 View
02 February 2020 2,956 3 View
I am facing problems lately with chemsketch freeware, I cannot copy and paste on word and powerpoint, nor edit chemsketch objects in these software. Can anyone advise me what can I do? I have...
29 January 2020 1,572 0 View
Hi All, I was wondering if anyone had used the chormotek gfp trap magenetic agarose bead kit for purification of GFP tagged nuclear proteins. I am attempting to purify a gfp-tagged protein...
15 January 2020 4,789 2 View
Hi, I was doing protein purification today and i observed that my niNTA beads turned brown after i bound my lysate with the beads. I eluted my protein but still the color of the beads were...
14 January 2020 7,541 4 View
I have a current conventional purification protocol on GSH-sepharose beads that I need to improve yield around 2.5-3 mg/mL, but also reproducibility and larger scale production. For this I was...
14 January 2020 6,437 4 View
Hi there, I´ve tried to purify a GST-tagged protein using IPTG and GST-Sepharose beads. Although I have used several bacterial volumes (50.100.200 ml), differents IPTG concentrations (0.5 and 0.2...
01 January 2020 4,067 4 View
01 January 2020 8,349 4 View
01 January 2020 8,799 2 View
Thioglycerol (3-mercapto-1,2-propanediol) is a sulfur-containing analogue of glycerol. This compound has been used as a reducing agent in protein purification as an alternative to other reducing...
01 January 2020 6,175 3 View
Hi, I am trying to purify a truncated part of protein that needs a proper post-translational modification, thus I use Flag-tagged mammalian construct and try purification with Flag agarose beads....
01 January 2020 9,538 3 View
I’m still a little new to this technique [of purifying his-tagged proteins]. I know there are protocols for recharging the columns but does the resin need to be removed before purifying the next...
31 December 2019 3,125 9 View
I need these bacterial strains for my research
28 December 2019 6,869 0 View
Can any one suggest me concentration of beta-mercaptoethanol (2-ME) or DTT to prevent cysteine dimerisation in protein purification at pH 8.0 . and stability of them in Tris Hcl buffer with...
17 December 2019 3,197 4 View
12 December 2019 6,202 4 View
12 December 2019 6,157 7 View
Does anyone have experience trying to work around background fluorescence in cyanobacteria? I'm currently trying to image a GFP fusion protein, but I didn't realize there was significant...
05 December 2019 6,000 3 View
I have a major problem that I have been facing for last few months. I have a protein which is His-tagged at the C-terminus. Its a fusion of 5 monomers and its weight is around 42 KDa. The...
15 November 2019 6,243 14 View
Hi All, I am currently working on a protein-miRNA binding assay. I am using FP as my assay system to see if the binding happens. Previous publications show the protein binds to the miRNA and my...
14 November 2019 8,836 2 View