I' new to purifying proteins and would love someone with experience
I have my protein now biotinylated (122 kDa in size).
I now want to purify the protein complex that binds to my super protein.
I liked magnetic beads however I tried Dynal M-280 Streptavidin beads and I am getting nothing compared to negative control....
I was conducting overnight binding at 4 degrees C in 50mM Tris 8.0, 150mM NaCl, 0.1% NP-40, 20% glycerol.
I was then thinking to now try sepharose beads.
Is this a good idea?
Any ideas why I may not be pulling down anything?
Your advise is greatly appreciated.