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
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 9,133 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 9,124 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 8,429 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 608 8 View
02 February 2020 6,219 2 View
02 February 2020 2,877 3 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 4,671 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 7,452 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 6,387 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 3,978 4 View
01 January 2020 8,256 4 View
01 January 2020 8,733 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 6,090 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 9,471 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 3,010 9 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 3,105 4 View
12 December 2019 6,113 4 View
12 December 2019 6,071 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 5,922 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 6,179 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 8,751 2 View
I am working on a membrane protein of size 70 kDa. It has a 6X-His Tag on C terminal. I am using 20mM Tris, 200 mM KCl, 10% glycerol, 5 mM beta-mercaptoethanol and 0.03% DDM for the protein...
11 November 2019 1,303 3 View
Dear All, I am using Anti-Flag magnetic beads for immunoprecipitation experiments. I want to perform elution under the non-denaturing conditions for further use of proteins as well as to reuse...
11 November 2019 4,866 7 View
11 November 2019 9,526 2 View