I want to detect a 3.5 kDa protein by western blotting and was wondering about the appropriate materials and conditions to follow. Please share your practical experiences.
Thanks!
Try using Tricine–SDS-PAGE.
https://www.nature.com/articles/nprot.2006.4
https://bio-protocol.org/bio101/e3093
Hello, Use a Tris-tricine gel, which is optimized for small peptides, and has short transfer time, bis-Tris SDS-PAGE, I will suggest also cast bis-tris gradient (15-20%) gels (Nu-PAGE).
https://openwetware.org/wiki/Sauer:bis-Tris_SDS-PAGE,_the_very_best
Good luck!
I see you already have some answers but Tricine gels are recommended along with the Tris-Tricine running buffer. If you are worried about the transfer on blots as well then try traditional methods? I worked with a 4KDa and even with a semi-dry transfer I managed to see my 4KDa monomer. But you can do traditional slow wet transfer to avoid losing your protein. Good luck!
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