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'd like to have an advice on which buffer and/or procedure I should use to maintain the phosphorylation status of a protein to be analysed through mass-spectrometry. I have to lysate breast...
11 May 2020 2,123 4 View
The proteins need to be enzymatically active, planning to use Taq. Polymerase for PCR reactions. I am looking to create a protein extract that can be used for diagnostic PCR. So the protein has to...
11 May 2020 7,273 12 View
Dear all, I'm looking for Cpf1 purification protocol and I found a protocol in which the authors added GABA to the buffers ( from equilibration till the eluition). I'd like to know if I can...
04 May 2020 7,757 6 View
In measuring the plasma HDL-c levels by a phosphotungstate-magnesium method, and the following enzymatic measurement of cholesterol in the supernatant, if I run out of the kit's standard...
01 May 2020 302 0 View
I have got multiple categorical variables which are all binary (yes/no). I would like to visualise 3-4 variables against one predictor variable in the same graph.
29 April 2020 1,648 0 View
We are expressing B-amyloid peptids and after purification we are having some problems with the aggregation of the samples. I'm wondering if there are optimal conditions of storage that dont...
08 March 2020 9,486 3 View
I'm extracting protein from highly hazardous bacterial samples. For samples like this, people commonly boil the bacteria prior to full lysis. Because the bacteria is boiled, do I still have to...
03 March 2020 6,074 3 View
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 900 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 9,154 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 9,134 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 8,449 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 620 8 View
02 February 2020 6,231 2 View
02 February 2020 2,892 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 1,533 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 4,688 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 7,478 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 6,395 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 4,000 4 View
01 January 2020 8,275 4 View
01 January 2020 8,749 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 6,110 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 9,480 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 3,032 9 View