Hello everyone out there?

09 September 2015 1 9K Report

I have a question regarding western blotting of large proteins particularly the clostridium difficile toxins. What SDS-PAGE (Stacking and Resolving gel) concentrations should i use? Even so, can i cleave these toxins and immunoblot another dormain such as the GTD, CROPS, etc? Please advise

Salma Teimoori

Dear Lincoln,

According to the information you gave, size of your protein is something about 300 kDa and the recommended gel percent is 6% resolving gel. You can also go for a gradient of 4-12% and see where you can find the protein more beautifully. About cleaving and blotting; of course you can do it as long as you have specific antibodies against the truncated ones.

Good luck!

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