Native molecular weight determination - using restricted SDS in PAGE (Oct/10/2010 )

I wish to determine molecular weight of the native protein (without dissociating the subunits). My protein will remain and migrate in intact form during the PAGE in the absence of SDS, but since there will be no uniform charge on the molecules, the separation will not occur only on the basis of size, hence I will not be able to determine the exact molecular weight. I am aware of other techniques such as gel filtration, blue-native PAGE which are more commonly used for this purpose. But since they require specialized chemicals and/or equipments, I was looking for a simpler method. While doing so, I came to know from here, that PAGE with restricted SDS can be used for native and subunit molecular weight determination simultaneously . I did one experiment and both native and denatured forms of my two test protein migrated the same distance which I did not expect. My next experiment will be doing the PAGE in the complete absence of SDS, where I wont be able to see the exact weight, but I can at least get an idea of what is happening. Meanwhile, does anybody has the experience with native molecular mass determination with this/similar method?
Thanks for your time

Hi Ram,
If you want to look for a native protein marker,
try look for NativeMark LC0725 from invitrogen
http://products.invitrogen.com/ivgn/product/LC0725?ICID=search-product

Hope this helps.

p/s: can you send me (pm) the article that you mentioned? I didn't subscribe to it. Thanks

adrian kohsf on Mon Oct 11 02:58:09 2010 said: