11 Questions 25 Answers 0 Followers
Questions related from Kiyohiro Takahasi
Hi I have expressed and purified 16 kDa protein and found this protein forms intermediate of monomer and dimer when it is highly concentrated. From some informations, it might be mediated by...
07 July 2018 2,509 2 View
Hi I have purified His-tagged protein. But after SDS-PAGE analysis I found a lot of protein seem to lost His-tag that is located at N-terminus and a lot of protein of interest might be lost at...
04 April 2018 7,584 4 View
Hi I have purified protein of interest. The sample was buffer exchanged to 20 mM HEPES pH 6.8, 200 mM NaCl and was concentrated by Amicon Ultra. The protein could be concentrated to at least 0.2...
12 December 2017 3,564 1 View
Hi I purified 14 kDa protein with Superdex 200 pg but the A280 nm of the target protein fraction was relatively low when it was compared to the amount of injected protein mixture. The injected...
10 October 2017 10,054 8 View
Hi I need suggestion about my current problem. I purified His-tagged protein using Ni-NTA resin and enough amount and purity of protein was observed on SDS-PAGE. But the protein was precipitated...
08 August 2017 9,439 5 View
I'm planning to make N-terminal split GFP fused recombinant proteins using E. Coli. So I want to make the expression vector with multi cloning site for target protein at the 3' of the split GPFs....
02 February 2016 8,958 4 View
Hi I am going to try E.Coli Rosetta 2 (DE3) pLyss strain for protein expression. I have read some papers and found pLys strain does not need sonication for lysing because of Lysozyme expression....
12 December 2015 7,278 3 View
I'm trying to purify membrane protein that is expressed in E. Coli. I have read several papers about lysing E. Coli Cells to purify target membrane protein. And found most of the researchers use...
03 March 2015 9,415 5 View
I'm trying to produce membrane protein using HEK293S cells. I have tested both transient expression with PEI and stable cell line and found transient expression can allow us to obtain higher...
02 February 2015 2,544 3 View
I'm trying to purify membrane protein from HEK293 cells for x-ray structural determination. I added the his-tag at the c-terminus of the protein, but the eluted protein from the resin contains...
12 December 2014 7,427 6 View
I'm trying to purify c-terminus his-tagged membrane protein that is stably expressed in HEK293S cells. I extracted membrane protein using detergent and confirmed the presence of the target protein...
11 November 2014 3,252 11 View
