Fabrication of carbohydrate chips and their use to probe protein-carbohydrate interactions.
Carbohydrate microarrays have received considerable attention as an advanced technology for the rapid analysis of carbohydrate-protein interactions. This protocol provides detailed procedures for the preparation of carbohydrate microarrays by immobilizing hydrazide-conjugated carbohydrates on epoxide-derivatized glass slides. In addition, we describe the use we make of these microarrays in glycomics research. Unlike other techniques that require large amounts of samples and long assay times, carbohydrate microarrays are used to carry out the rapid assessment of a number of carbohydrate-recognition events with tiny amounts of carbohydrate samples. Furthermore, the microarray technology is also utilized for the rapid assay of enzyme activities. We are able to routinely prepare carbohydrate microarrays within 12 h by using hydrazide-conjugated carbohydrates and apply these microarrays for the studies of glycan-protein interactions within 8 h.
Park
Sungjin
S
Department of Chemistry, Yonsei University, Seoul 120-749, Korea.
Lee
Myung-Ryul
MR
Shin
Injae
I
D001665:Binding Sites; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000138:chemical synthesis*; D002352:Carrier Proteins / Q000032:analysis*; D004852:Epoxy Compounds / Q000737:chemistry; D006834:Hydrazines / Q000737:chemistry; D046228:Microarray Analysis / Q000379:methods*; D015335:Molecular Probes / Q000138:chemical synthesis*
D002241:Carbohydrates; D002352:Carrier Proteins; D004852:Epoxy Compounds; D006834:Hydrazines; D015335:Molecular Probes; C029424:hydrazine