try codon optimisation
use Rosetta 2 strains
fuse to MBP
I expressed a yeast protein of ~100 KDa, expression was low until I fused it
to MBp (resulting protein ~ 150 KDa)
Hello everyone,
Just want to say thanks for your great ideas and time to reply my question.
Hope I will solve my problem soon
Kien
Hello everyone,
I am expressing a 100 KDa eukaryotic membrane protein in E coli. The protein
is fused to 6His-MBP in the N terminus and the resulting mass is ~ 150 KDa.
However, the protein get severely degraded so after putting through a Ni-NTA
column, the protein came out with a lot of
Dear all,
I am wondering if anyone is working on Blue Natve PAGE or other alternate
methods. I need some help to troubleshoot my Native PAGE experiments. It
will be great if anyone can help me in this regard. I am working on membrane
proteins which have pI around less than 7. i need to run
Hello everyone...
I have a protein solubilised in 1% LDAO, 50 mM Phosphate pH 7.5, 150 mM
NaCl. When I mix it with laemmli buffer and heat at 95C, 3 mins for
SDS-PAGE, it aggregates (as in when you mix GuHCl with SDS) and can't be
loaded onto the gel.
Does anyone know why it happens? Thanks a
