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Electrophoresis Buffer
Electrophoresis buffer is a critical component of nucleic acid and protein gel electrophoresis systems, serving as a conductor in the electric field and maintaining a stable pH. Its composition and ionic strength influence the electrophoretic mobility of analytes and must not react chemically with samples to avoid altering their physicochemical properties or causing loss of biological activity. EDTA in the buffer chelates divalent cations such as Mg²⁺, preventing activation of nucleases during electrophoresis and avoiding precipitation between Mg²⁺ and nucleic acids.
Figure 1. Chemical structure of EDTA
Native-PAGE (native polyacrylamide gel electrophoresis) and SDS-PAGE (sodium dodecyl sulfate denaturing polyacrylamide gel electrophoresis) both utilize polyacrylamide gels for protein separation; however, they differ in separation principles, objectives, and sample states.
Native-PAGE (non-denaturing electrophoresis)
✦ No SDS or denaturants are added, and reducing agents (e.g., β-mercaptoethanol, DTT) are either omitted or minimally used.
✦ Proteins maintain their native conformation, preserving secondary, tertiary, and quaternary structures.
✦ Separation depends on a combination of protein net charge, size, and shape.
✦ Enables detection of protein complexes, conformational states, and biological activity.
✦ Commonly used to study protein interactions, complex composition, and function.
SDS-PAGE (denaturing electrophoresis)
✦ SDS is added to the sample, often combined with reducing agents and heat treatment.
✦ SDS denatures proteins by fully unfolding them into linear chains and imparts a uniform negative charge.
✦ Separation is based solely on protein molecular weight.
✦ Enables precise determination of protein molecular weight.
✦ Commonly used for assessing protein expression levels, purity, and molecular weight analysis.
Native-PAGE (non-denaturing electrophoresis) products
ID | Product Name | Introduction |
Tris-MOPS-Native Running Buffer (Powder) | Suitable for electrophoresis buffer of Bis-Tris system gel | |
Tris-Hepes-Native Running Buffer (Powder) | Suitable for electrophoresis buffer of Hepes-Tris system gel | |
Tris-MES-Native Running Buffer (Powder) | Suitable for electrophoresis buffer of Bis-Tris system gel | |
Tris-Acetate-Native Running Buffer (Powder) | Adapted to Tris-Acetate system gel, commonly used for separating macromolecular proteins | |
Tris-Glycine-Native Running Buffer (Powder) | Suitable for electrophoresis buffer of Tris-Gly system gel; Not suitable for gels of buffer systems such as Hepes and Bis-tris |
SDS-PAGE (denaturing electrophoresis) products
ID | Product Name | Introduction |
Tris-MOPS-SDS Running Buffer (Powder) | Suitable for electrophoresis buffer of Bis-Tris system gel | |
Tris-Hepes-SDS Running Buffer (Powder) | Suitable for electrophoresis buffer of Hepes-Tris system gel | |
Tris-MES-SDS Running Buffer (Powder) | Suitable for electrophoresis buffer of Bis-Tris system gel | |
Tris-Acetate-SDS Running Buffer (Powder) | Adapted to Tris-Acetate system gel, commonly used for separating macromolecular proteins | |
Tris-Glycine-SDS Running Buffer (Powder) | Suitable for electrophoresis buffer of Tris-Gly system gel; Not suitable for gels of buffer systems such as Hepes and Bis-tris |
Aladdin: https://www.aladdinsci.com/
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