Help! I need to run collagen on SDS gel - (May/17/2007 )
Could someone plese help me?! My boss wants to run collagen extracted from the tissue (heart) on SDS gel. I have no idea where to start. How do I extract collagen? What standard do I run?
I have extracted collagen form calf tendon (collagen rich) previously which involved dissolving the collagen in acetic acid (0.2 M) for approx 2 days at 4 degrees with stirring, you then precipitate the collagen by neutralising with NaOH
not too sure how you would get it purified from the homogenate of heart
a standard 5% separarting gel should be fine with some high range molecular weight markers
intact collagen should run as 3 distinct bands.
you could run some from sigma alongside though the stuff i have bought from them in the past has only been good for throwing in the bin !!
hope this helps
I am planning to run collagen on a SDS gel and wanted to know if you figured out the protocol to use. I also need to know what is the best way to extract the collagen from my tissue samples that are mouse heart. I hope you got good results that may be useful for me