Protein Separation Myths
 | It's about time old electrophoresis myths were busted. Don't get left behind. Bring your separation tools up to date today with Novex® gels. Protein Separation Myths: 1. Gel chemistry will always modify my protein 2. NuPAGE® gels take too long to run |
MYTH: Gel Chemistry Will Always Modify my Protein
FACT: NuPAGE® Gel Chemistry Actually Preserves Your Protein
More Neutral pH is the KeyThe fundamental shortcoming of the tris-glycine gel system is protein degradation during sample preparation. When using a tris-glycine sample buffer prepared at pH 6.8, samples are heated within the buffer, resulting in a pH decrease due to the negative temperature coefficient of tris-glycine. This ultimately results in protein hydrolysis. NuPAGE® gel chemistry preserves protein sample integrity by maintaining a higher sample buffer pH (Figure 1). The pH of the NuPAGE® sample buffer does not drop below pH 8 (even when heated to 70°C), so the protein sample does not degrade during sample preparation. | Meet the inventor of the NuPAGE® System and learn about preserving your protein during electrophoresis. |
| Figure 1. See the difference with NuPAGE® gels. (A) The NuPAGE® gel run using 1X MES buffer. (B) A tris-glycine gel run using tris-glycine/SDS buffer. This figure demonstrates that NuPAGE® gels will produce sharp, intact bands that represent unhydrolyzed protein when compared to tris-glycine gels, particularly evidenced in lanes 3, 5, 7, 8 and 9. Samples loaded: (1) Mark12™ Unstained Standard, 10 μL; (2) rat liver lysate, 10 μg; (3) lysozyme, 6 μg; (4) Bio-Rad® Broad Range Standard, 1:50 dilution; (5) BSA, 6 μg; (6) MagicMark™ XP Standard, 10 μL; (7) human IgG, 6 μg; (8) human IgM, 6 μg; (9) E. coli lysate, 10 μg; (10) Novex® Sharp Unstained Standard, 10 μL. |
MYTH: NuPAGE® Gels Take Too Long to Run
FACT: NuPAGE® Gels Can Be Run in 24 Min at 250V
Standard Run ProtocolTo obtain efficient separation, traditional Laemmli gels can take as long as 90 min to run. In contrast, the standard protocol for NuPAGE® gels requires 35 min at 200V. Moreover, our accelerated protocol can typically produce efficient separation in just 24 min at 250V (Figure 2).
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Observe a Typical Run |
Figure 2. Accelerate run time without sacrificing performance. NuPAGE® samples loaded: (1) Mark12™ Unstained Standard, 10 μL; (2) rat liver lysate, 10 μg; (3) lysozyme, 6 μg; (4) Bio-Rad® Broad Range Standard, 1:50 dilution; (5) BSA, 6 μg; (6) MagicMark™ XP Standard, 10 μL; (7) human IgG, 6 μg; (8) human IgM, 6 μg; (9) E. coli lysate, 10 μg; (10) Sharp Unstained Standard, 10 μL.MYTH: Precast Gels are too Expensive
FACT: Using Novex® Precast Gels Helps Save Not Only Time and Precious Samples, but Also Money
Traditional gel separation hasn't changed much over the last 30 years. Using Novex® precast gels offers the following significant advantages:
Learn more about Novex® Precast Gels Learn more about our gel casting essentials
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