Superior resolution of large molecular weight proteins
Invitrogen NuPAGE Tris-Acetate Gels simulate the denaturing or the native conditions of the traditional Laemmli system. A unique buffer formulation helps to maintain a low operating pH during electrophoresis, resulting in superior resolution of proteins compared to traditional tris-glycine SDS-PAGE gels.
Reliable separation and transfer of high molecular weight (HMW) proteins is a common challenge for life science researchers. Choosing the right gel is a key factor in the successful transfer of HMW proteins. A popular general-use gel is a 4–20% tris-glycine gel, which can effectively separate a mixed range of proteins. However, HMW proteins will be compressed into a narrow region at the top of the gel. A better option for HMW proteins is a tris-acetate gel or a low percentage non-gradient tris-glycine or bis-tris gel. When specially targeting HMW proteins, optimal transfer can be achieved with a tris-acetate gel.
Tris-acetate gels maintain a neutral pH and separate HMW proteins with higher resolution than bis-tris or tris-glycine gels. Comparison of HMW protein separation using different gel chemistries and gradients shows best separation and resolution of HMW proteins using a 3–8% tris-acetate gel. This increased resolution leads to increased transfer efficiencies and higher sensitivity.
Sample preparation is a crucial step in successful protein analysis. The pH of the traditional Laemmli-style sample buffer changes from 6.8 to 5.2 when heated at 100°C. This lower pH is known to induce aspartyl-prolyl (Asp-Pro) peptide bond cleavage, which leads to protein degradation. By maintaining a >7.0 pH environment, the Invitrogen NuPAGE LDS Sample Buffer preserves protein integrity by minimizing this Asp-Pro cleavage. Sample integrity is maintained throughout electrophoresis with the Invitrogen NuPAGE system; protein degradation is seen in samples prepared with Laemmli (tris-glycine) sample buffer. The Invitrogen NuPAGE Antioxidant running buffer additive greatly minimizes protein oxidation during electrophoresis and keeps reduced protein bands sharp and clear.
For Research Use Only. Not for use in diagnostic procedures.