Shreffler: SDS-PAGE
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Overview
From wikipedia: "SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis, describes a technique widely used in biochemistry, forensics, genetics and molecular biology to separate proteins according to their electrophoretic mobility (a function of the length of a polypeptide chain and its charge). In most proteins, the binding of SDS to the polypeptide chain imparts an even distribution of charge per unit mass, thereby resulting in a fractionation by approximate size during electrophoresis."
We use the Invitrogen NuPAGE system.
Materials
- Appropriate gelbox/voltage source
- NuPAGE 4-12% BIS-TRIS Gel (Invitrogen NP0322BOX)
- NuPAGE MES SDS Running Buffer (20x) (Invitrogen NP0002)
- NuPAGE SimplyBlue SafeStain (Invitrogen LC6060)
- NuPAGE LDS Sample Buffer (4X) (Invitrogen NP0007)
- NuPAGE Antioxidant (Invitrogen NP0005)
- NuPAGE Reducing Agent (Invitrogen NP0004)
Procedure
Preparing Samples
1. We want to Add 5 μL 4X Reducing Buffer (RB) to 15μL protein solution. Vortex sample to mix. Heat samples at 70 °C for 10 mins. Spin samples.
Discussion
Contact
- Who has experience with this protocol?
- Email Alex Ma through OpenWetWare
