ProfilerPro Glycan Profiling User Guide - PerkinElmer

ProfilerPro Glycan Profiling Kit, Ver 2, User GuideIdentifying peaks by co­migration with known Glycan StandardsIt is sometimes interesting to compare the migration time of peaks observed in your samples to the migrationtime of known glycan standards, to help identify the peaks in your samples. When running glycan standards it isimportant that the buffer the standards are presented in is closely matched to the buffer your samples arepresented in. If the buffers are not well matched, the migration time of the standards may not correspond to themigration time of your sample. A simple method to achieve matched buffers is to label your standards using thereagents from the Glycan Release and Labeling kit, similar to the method used to prepare your test samples,but eliminating the denaturing and digestion incubation steps as shown in the diagram below. Any dilutions ofthe standards prior to addition in the Denaturing Plate should be done with molecular grade water.No incubation.No incubation.Label for2hrs at 55C.8μL standardsampleTransfer ~11μLof denature.Transfer 8μLof digest.Reconstituteand run.Denaturing Plate PNGase F Plate Dye Plate LabChip GXIILMG0G0fG2Man5G1f’’G1fG2fExample showing a mix of glycan standards compared to an IgG sample, run in well matched buffers.Blue = IgG, Red = Mix of six glycan standards (Man5, G0, G0f, G1f, G2, G2f). The standards were runat about 50 ng/µl concentration.Caliper Life Sciences Page 9 of 21 PN: 45070968 Elm StreetHopkinton, MA 01748-16681-877-LABCHIP (522-2447)