Western blot for muscle - (Mar/22/2010 )
How's it going everyone? Just registered here, this looks like quite an extensive and valuable resource for lab work. My question is about Western blots for muscle. I work in a neuro lab so our Western protocol is great for neurons. But recently I've been working with NMJs and we want to look at proteins in muscle specifically (both skeletal and smooth). Our Western protocol isn't great for muscle so I'm wondering if anyone has a tried and true protocol (that is, buffer recipes and anything else of note). My primary search of the literature was a bit scattered so I'm hoping I can clear it up here before leaping into the literature more.
I don't work with muscle, but you will need a pretty strong homogenisation process to grind the muscle up - either use a polytron like thing or snap freeze and grind in LN2 or dry ice.
bob1 on Mar 22 2010, 08:24 PM said:
we used to use a meat grinder for the first step.