title: General protein-protein cross-linking.
abstract: This protocol describes a general protein-to-protein cross-linking procedure using the water-soluble amine-reactive homobifunctional BS(3) (bis[sulfosuccinimidyl] suberate); however, the protocol can be easily adapted using other cross-linkers of similar properties. BS(3) is composed of two sulfo-NHS ester groups and an 11.4 Å linker. Sulfo-NHS ester groups react with primary amines in slightly alkaline conditions (pH 7.2-8.5) and yield stable amide bonds. The reaction releases N-hydroxysuccinimide (see an application of NHS esters on Labeling a protein with fluorophores using NHS ester derivitization).
authors.affiliation: Thermo Fisher Scientific, Rockford, IL, USA. Electronic address: alice.schaffer@thermofisher.com.
authors.forename: Alice
authors.initials: A
authors.lastname: Alegria-Schaffer
mesh_terms: D002623:Chemistry Techniques, Analytical; D003432:Cross-Linking Reagents / Q000737:chemistry; D006868:Hydrolysis; D011506:Proteins / Q000737:chemistry*; D013388:Succinimides / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins; D013388:Succinimides; C035760:bis(sulfosuccinimidyl)suberate
keywords: Amount of BS(3); Hydrolysis of NHS ester; Moisture condensation; Phosphate-buffered saline (PBS); Protein cross-linking