Native PAGE

Native PAGE separations are run in non-denaturing conditions. Detergents are used only to the extent that they are necessary to lyse lipid membranes in the cell. Complexes remain—for the most part—associated and folded as they would be in the cell. One downside, however, is that complexes may not separate cleanly or predictably, since they cannot move through the polyacrylamide gel as quickly as individual, denatured proteins.

Unlike denaturing methods, such as SDS-PAGE, native gel electrophoresis does not use a charged denaturing agent. The molecules being separated (usually proteins or nucleic acids) therefore differ not only in molecular mass and intrinsic charge, but also the cross-sectional area, and thus experience different electrophoretic forces dependent on the shape of the overall structure. Since the proteins remain in the native state they may be visualised not only by general protein staining reagents but also by specific enzyme-linked staining.

[cheetengchay]

why the preparation of sample was using beta-Mercaptoethanol? Isn't native PAGE should not used any denaturing agent such as SDS and Mercaptoethanol??

[Ramk]

Good video and useful to the researchers...nice work...

[adva]

Water has to be removed from above resolving gel

[gasperkosmac]

hey does this protocol work? I tried one before which had a lot more APS in it and the gel got all rigid & crumbly, suffice to say it failed lol :)

[Crispercas]

No mention of the PH and the PI, this is a native PAGE not an SDS PAGE!

[Crispercas]

This is definitely south of India, I can bet a 1000 dollars, they walk like this all the time :P

[Deby_M]

WTH... The person in 2.28 min is walking in naked feet in the lab