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• W2082027989 abstract "A two-step zero-length crosslinking procedure for studying protein-protein complexes has been developed. One component of a complex is briefly incubated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) in the presence of N-hydroxysuccinimide resulting in the conversion of some of the protein carboxyls into succinimidyl esters. The reaction is stopped by addition of β-mercaptoethanol and other interacting proteins are then added. Crosslinking arises from substitution of lysine ϵ-amino groups of these proteins for the succinimidyl moieties during a 1- to 2-h incubation period. The advantage of this method versus one-step zero-length crosslinking is that only one component of the complex is exposed to the crosslinker, which eliminates complications arising from the formation of crosslinks among several proteins of a multicomponent complex. Furthermore, crosslinks can be formed even in the presence of reagents, such as dithiothreitol and EDTA, that would interfere with direct crosslinking with EDC." @default.
• W2082027989 created "2016-06-24" @default.
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• W2082027989 date "1990-02-01" @default.
• W2082027989 modified "2023-10-18" @default.
• W2082027989 title "Zero-length crosslinking procedure with the use of active esters" @default.
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• W2082027989 doi "https://doi.org/10.1016/0003-2697(90)90267-d" @default.
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