SDS-PAGE Identification of Target Proteins

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is one of the most commonly used high-resolution analytical separation electrophoresis techniques. Electrophoresis involves the movement of charged molecules through a matrix under the influence of an electric field. In this technique, factors affecting the mobility of charged molecules include their relative molecular weight, charge, and structure. SDS-PAGE uses an anionic detergent to initially denature the sample, imparting a negative charge proportional to the molecular weight of the molecules being tested. Combined with polyacrylamide gel, this technique eliminates the effects of structure and charge, allowing substances to be separated solely based on molecular weight differences. It is commonly used in high-resolution separation experiments of large molecules, including complex DNA, RNA, and proteins.

When identifying proteins using SDS-PAGE, both single proteins and protein mixtures can be analyzed. Proteins in the sample migrate according to their relative molecular weight, and after electrophoresis, proteins of different molecular weights are separated into distinct bands. For single protein samples, it can be used for molecular weight identification and to verify if other protein impurities are present. For protein mixtures, SDS-PAGE electrophoresis can separate different components in the mixture and identify their relative molecular weights. Additionally, the width of the protein bands after electrophoresis can provide a rough estimate of their abundance.

Biotech company Biotai Park uses the Bio-Rad Mini-PROTEAN® Tetra Gel System and Thermo's latest Obitrap Fusion Lumos mass spectrometer, combined with nanoLC-MS/MS nanoliter chromatography, to provide SDS-PAGE and mass spectrometry-basedprotein identificationservice packages, which can quickly and accurately identify the relative molecular weight, purity, structure, amino acid composition, and sequence of various protein samples. Additionally, Biotai Park offers customized analysis services to meet different experimental needs. Free consultations are welcome.

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Molecular weight determination
Identification of protein spots, bands, and IP sample proteins
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