Western Blot problems

Laurence Pelletier pelletl at IRCM.UMontreal.CA
Sat Dec 28 17:21:50 EST 1996


I am currently having problems western blotting total yeast extracts i
prpare using glass beads. The protocol i use is quite simple: you vortex 5
times one minute at 4 degres, spin down one minute at 1000 rpm in a
tabletop microfuge and collect the supernatant. I then submit 25
micrograms of total protein to western blotting using BM Chemiluminescence
Kit using their protocol. 

Here's the problem: Most of the time (9 out of 10) the blots are full of
streaks only in the lanes that contain samples (including molecular weight
markers). Once in a while, i have near perfect blots (just to stay
humble). 

Any ideas?

Happy New Year

Laurence Pelletier
pelletl at IRCM.Umontreal.CA





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