2D Blue Native/SDS-PAGE Complex Analysis Service

In biological membranes, many proteins form complexes. Biotech company Biotai can perform global analysis of the subunits of these protein complexes using 2D Blue Native/SDS-PAGE. In 2D BN/SDS-PAGE analysis, samples are first separated by blue native polyacrylamide gel electrophoresis (BN-PAGE) in the first dimension, and then by SDS-PAGE in the second dimension.SDS-PAGE) for one-dimensional separation, followed by two-dimensional separation using SDS-PAGE.

2D Blue Native SDS-PAGE Complex Analysis Service

Protein complexes can organize and maintain cellular and organelle functions across all complex temporal and spatial levels, including cell development and division, transcription and translation, respiration and photosynthesis, transport, and metabolism. Protein complexes may exhibit partial and temporary variations, and demonstrate different stabilities, making identification and analysis challenging. BN-PAGE is a technique used for the high-resolution separation of protein complexes. BN-PAGE can be used to determine the size, relative abundance, and subunit composition analysis of protein complexes.

In 2D BN-PAGE analysis, protein or protein complex samples are separated under native conditions in 1D BN-PAGE. Coomassie Brilliant Blue dye, which carries a negative charge, non-specifically binds to all proteins. Coomassie Brilliant Blue is not used as a detergent, thereby preserving the structure of protein complexes. Additionally, the binding of Coomassie Brilliant Blue reduces the tendency for protein aggregation during electrophoresis. Therefore, the electrophoretic mobility of the samples depends on the negative charge of the bound Coomassie Brilliant Blue as well as the size and shape of the complexes. After separation by BN-PAGE, the protein and protein complex samples are denatured in the gel with SDS before being further separated by 2D SDS-PAGE gel electrophoresis.

The combination of BN-PAGE and SDS-PAGE, due to the gradient gel in the first dimension and the linear gel in the second dimension, results in monomeric proteins migrating along a hyperbolic diagonal, while the constituent components of protein complexes are found below this diagonal. Proteins representing the same protein complex subunits can be found in a vertical line in the second dimension, while multiple spots of the same protein in a horizontal line indicate that the protein is simultaneously present in several different protein complexes.
2D BN/SDS-PAGE analysis can provide information about the size, quantity, protein composition, stoichiometry, and relative abundance of samples, making it a useful tool for studying protein complexes.