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Advantages of the NativePAGE Bis-Tris gel system include:
•Broad molecular weightAnalytical range: 15–10,000 kDa
• Neutral pH separation, Able to better maintain the nondenatured state of protein complexes
• All proteins with different isoelectric points (pI) in separable gels
•Able to maintain the nondenatured conformation Analyze membrane-protein complexes
• Can obtain higher resolution than traditional tris-glycine native electrophoresis
NativePAGE Bis -How Tris gels work
In SDS-PAGE, SDS acts as a charge transfer molecule, which gives the protein a net negative charge, thereby denaturing the protein and migrating toward the anode. BN PAGE uses Coomassie G-250 dye as a charge transfer molecule, which attaches to the protein to give it a net negative charge while maintaining the protein in its non-denatured, native state. The NativePAGE Bis-Tris gel system has a near-neutral pH environment, which can provide greater stability to the protein and gel matrix, allowing for highly sensitive analysis of non-denatured membrane-protein complexes, compared to traditional Tris-glycine precast The gel system provides excellent band resolution.
We recommend using NuPAGE Transfer Buffer for traditional wet transfer and PVDF to transfer proteins to the membrane. NuPAGE Transfer Buffer maintains the neutral pH environment established during electrophoresis. Nitrocellulose membranes are incompatible with blotting membranes because nitrocellulose membranes bind Coomassie G-250 dye very tightly. Alternatively, use the Invitrogen Power Blotter (PB0013) for fast semi-dry transfer or the iBlot 2 Gel Blotter with PVDF membrane (IB21001) for fast dry transfer.