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Cross-linking refers to the process of chemically bonding two or more molecules through a covalent bond. The crosslinker comprises a reactive end which is reactive with specific functional groups located on proteins or other molecules, such as primary amines and sulfhydryl groups. Multiple chemical groups on proteins and polypeptides can be used for cross-linking, so proteins and polypeptides can be used as targets for coupling in studies using cross-linking methods.
Cross-linkers are used to determine the relationship between adjacent proteins as well as ligand-receptor interactions. Photo-reactive crosslinkers are widely used for nonspecific bioconjugation. While numerous options exist, the two most common photo-reactive chemical groups are diazirines and aryl-azides. Photo cross linkers are used for the study of protein-protein or protein-oligonucleotide interactions in living cells. They are typically used in the following manner:
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