Functional Glycomics, 1st Edition by Minoru Fukuda

By Minoru Fukuda

During this three quantity assortment targeting glycomics, readers will relish how such discoveries have been made and the way such equipment could be utilized for readers' personal learn effortsEach bankruptcy has been designed in order that adequate medical historical past might be given in every one bankruptcy for additional improvement of equipment by way of readers themselves. worthwhile for all degrees of scientists ranging from the final years of schools, graduate scholars, postdoctoral fellows to professors and to all degrees of scientists in examine institutes together with undefined.

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Individual clones were picked and analyzed. 2. Lectin blot and lectin staining analysis Cells were harvested, rinsed with phosphate-buffered saline (PBS), and lysed with 1% Triton X-100 in PBS. Cell lysates containing 30 mg of protein were boiled in SDS sample buffer with b-mercaptoethanol, loaded on SDSPAGE, and then transferred onto a PVDF membrane. Then the membrane was treated with 25 mM H2SO4 at 80  C for 60 min to remove sialic acid residues. After being blocked with 5% bovine serum albumin (BSA), the membrane was incubated with HRP-lectin for 2 h at room temperature.

Dual luciferase assay Cells transiently transfected with pGL3-b1,4GalT V promoter and pRLCMV were washed and lysed in 100 ml of passive lysis buffer (Promega). Firefly luciferase and Renilla luciferase activities were measured with 5 ml of cells lysate using the Dual-Luciferase Reporter assay system (Promega) in a luminometer. ‘‘Relative activity’’ was defined as the ratio of firefly luciferase activity to Renilla luciferase activity and was calculated by dividing 8 Jianhai Jiang and Jianxin Gu luminescence intensity obtained in the assay for firefly luciferase by that obtained for Renilla luciferase.

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