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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...
18 August 2016 2,881 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,...
18 May 2015 2,095 6 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...
16 March 2015 3,740 4 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...
09 December 2014 1,362 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...
22 November 2014 2,352 6 View
