Hi,everyone
I'm doing experiments on exosome. And Western blot is a well established method to detect exosome. The tetraspanins CD9, CD63 and CD81 are commonly used for exosome detection. I have noticed that exosomal protein are run on non-reducing gel in some papers. Can I use reducing SDS loading buffer and reducing gel? And could you tell why?
I am very grateful for your opinions!
Dear Hua Gai,
you are right. For detection of CD9, CD63 or Cd81 you must not use reducing conditions. The disulphide bonds in the LEL are important for 3D structure and conformational epitopes are recognized by the antibodies.
Best regards, Luise
Dear Luise ,
Thank you very much!
Reducing conditions can reduce disulphide bonds so that proteins can be better separated by size in SDS-PAGE gels. As you said, the disulphide bonds are important for 3D structure and conformational epitopes are recognized by the antibodies of CD9, CD63 and CD81. I still have a little doubt about this problem. Why are conformational epitopes important for antibodies of the tetraspanins, but not other proteins？
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