7 Questions 22 Answers 0 Followers
Questions related from Zhenning Tan
I have been struggling with purification of a highly basic protein. This protein is His-tagged. We get pretty good purity from Ni column. The pooled Ni column fractions is in 500 mM NaCl, ~ 200 mM...
08 August 2016 4,460 9 View
Currently, I am performing cell fractionation to separate cytoplasmic and nuclear fractions to quantify the localization of my protein. Right now, I am test the fractionation method. I am using...
08 August 2015 1,335 8 View
I have used midi prep kit to purify plasmids and found that the transfection efficiency for HepG2 cells not very high, about 30%. I am using Turbofect from life technologies. My questions are, 1,...
05 May 2015 5,086 7 View
I have been looking at colocalization of two proteins in cells with fluoresence microscope. I take three channels of pictures for each slide ( DAPI, green and red). All original files are tiff and...
04 April 2015 1,127 7 View
I am cloning a vector from a 6 kb plasmid. I started with 10 ug plasmid. I did double digestion to remove 2kb fragment and then used phosphotase to treat the vector. After steps of digestion and...
03 March 2015 5,020 5 View
Does anyone have any idea how to use affinity column to purify Avitagged protein? This protein is biotinylated in vivo by a biotin ligase? Can I bind it to Strepavidin beads and elute it with free...
12 December 2014 2,957 8 View
I have been doing immunostaining of cultured cells. The current issue is that I don't see any positive stain. One thing that worries me is the fixation and permeabilization step. Is it possible...
11 November 2014 1,628 7 View
