Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 15)

Section 4
Native PAGE
4.1 Introduction
Ready Gel Tris-HCl gels are made without SDS, allowing separation of protein in their native conformation.
The nonreducing and nondenaturing environment of native PAGE allows the detection of biological activity
and can improve antibody detection. Native PAGE can also be used to resolve multiple protein bands where
molecular mass separation by SDS-PAGE would reveal only one.
Native PAGE uses the same discontinuous chloride and glycine ion fronts as SDS-PAGE to form moving
boundaries that stack and then separate polypeptides by charge to mass ratio. Proteins are prepared in a
nonreducing nondenaturing sample buffer, which maintains the proteins’ secondary structure and native
charge density. Native PAGE is not suitable for accurate molecular weight determination due to the variability
of charge to mass ratio among different proteins.
4.2 Ready Gel Tris-HCl Gel Composition
Gel buffer 0.375 M Tris-HCl, pH 8.8
Cross-linker 2.6% C
Stacking gel 4% T, 2.6% C
Storage buffer 0.375 M Tris-HCl, pH 8.8, NaN
3
Shelf life 12 weeks from the date of manufacture
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