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 exploring potential chemical based approaches for cell cycle synchronization and would like to hear from anyone with experience on this technique what they believe to be the most applicable...
26 April 2021 2,760 3 View
Hi guys, I wondered anyone know how to disassemble ATP synthase complex in vitro by any drug or other method? Thanks,
14 April 2021 9,311 4 View
For my research, I need to collect the information regarding the approximate quantity of- 1. The number of Cytochrome c oxidase (COX) are present in a single mitochondria 2. The number of...
11 March 2021 7,004 0 View
I measured the OD600 for C. crescentus wild type and mutants strains every 60 minutes, and I need to calculate the doubling time for each strain to see the effect of mutations on doubling time....
26 February 2021 2,289 2 View
We are about to recombinantly express a couple of membrane proteins in E. coli and then purify them using detergent-containing buffers. We'd like to first screen a panel of detergents on a small...
23 February 2021 5,016 3 View
I am working on a project where I have to purify some proteins that are tagged with a 6x His tag. Since my proteins were produced by bacteria, I purchased the HOOKTM 6X His Protein Purification...
10 February 2021 7,275 4 View
UV-Visible Spectrophotometric evaluation of crude phycobiliproteins shows absorption peaks corresponding to both phycocyanin and phycoerythrin. so, how can I separate both phycocyanin and...
05 February 2021 8,302 2 View
Hello, I currently have sliced and punched mouse brain tissue using the cryostat and I wish to analyze it for the concentrations of different dopamine receptors. We initially desired to use...
02 January 2021 9,669 2 View
Hi everybody, Does anyone have a good protocol for protein purification using AKTA? I have tried to purify two proteins but after several attempts it was not successful. So, I really appreciate...
29 December 2020 5,574 15 View
Hi everyone, I am doing protein expression via E.coli. I am required to lyse the e.coli cells via sonication to obtain cell lysate. I normally sonicate 1L cell pellet suspended in salt buffer at...
23 December 2020 9,930 7 View
Hi, I am currently working with BL21(DE3)PLySs and PRSET A vector. Out lab frequently transformation and culture E Coli for protein purification. But recently, even though there is colony after...
14 December 2020 9,505 9 View
I have flash-frozen brains and am looking to extract RNA for RNA-seq, and protein for western blotting. I've seen other questions related to this...
11 December 2020 6,531 0 View
Based on this paper The P1 ' specificity of tobacco etch virus protease ,TEV protease can cleave between the Gln and several amino-acids (besides Gly/Ser) with acceptable efficiency in its...
27 November 2020 6,234 4 View
During protein purification, a protease inhibitor is added to the lysate (10ul/mL of lysis buffer). Why is this amount of inhibitor desirable? What is the potential danger to the protein if a...
24 November 2020 8,430 7 View
pure protein , methods to puried , characterized pure protein
17 November 2020 7,579 3 View
I have a protein labeled with a fluorescent dye (fluorescein) and I am hoping to get a quick measure of purity using TLC after dialysis. TLC is quick and easy so I am hoping to start there. What...
16 November 2020 7,522 3 View
I purchased this Merck millipore's DURAPORE PVDF 0.45 micron WH PL 13 mm hydrophobic filter instead of 0.45 um syringe filter. Now I am wondering if it is suitable to use it for purifying my...
10 November 2020 1,296 5 View
I am going to extract proteins, and for that, I will use a protease inhibitor cocktail and Phospho-Stop (from Sigma). For protease inhibitor stock solution (2x conc.), it is mentioned through the...
27 October 2020 4,528 3 View
Hello all, I'm currently working with undergraduate student-researchers (there are no graduate students at my university) to purify and characterize the drug targetability of glucokinase from pig...
26 October 2020 3,929 7 View
Hello, I am interested in producing purified Cas9 in my lab. But we are not dealing with proteins so we don't have proper settings for protein purification. It seems a step using chromatography is...
26 October 2020 10,118 3 View
We have always used Probond (Invitrogen) but Promega resin is cheaper. I was wondering how the Promega resin performs, to see if it is worth to change brands. Thank you!!
19 October 2020 4,637 3 View
Hello, I have a question for old school scientists. I read in an old book about protein purification about the vacuum dialysis method for volume reduction and buffer change. When a sample is...
13 October 2020 687 8 View
I'm currently doing protein purification. my protein size is around 42kDa, however, my protein is attached with fusion tag which are Trx-tag and His-tag at the N terminal. I did optimize my buffer...
13 October 2020 2,272 3 View
We are trying to save up on reagents and heat treatment is a promising protocol, but there are limitations with the thermostability of strand displacing DNA polymerases. Does anyone know of a...
07 October 2020 4,900 4 View