Journal: eLife

Article Title: A robust method for particulate detection of a genetic tag for 3D electron microscopy

doi: 10.7554/eLife.64630

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , CTP , Chem-Impex , 00095 , .

Techniques: Transfection, Construct, Recombinant, Sequencing, In Vitro, Labeling, Electron Microscopy, Inhibition, Purification, Software

Journal: Archives of biochemistry and biophysics

Article Title: INCORPORATION OF PHOSPHATE INTO GLYCOGEN BY GLYCOGEN SYNTHASE

doi: 10.1016/j.abb.2016.03.020

Figure Lengend Snippet: Effect of treating 32P-labeled glycogen with laforin or glucosidases. A Glycogen was labeled by incubation with 5 μM [β-32P]UDP-glucose (lower panel) or [U-14C]glucose (upper panel) and yeast glycogen synthase (2 μg/ml) for 30 min. Glycogen was precipitated with ethanol, dissolved in buffer and treated with α-glucosidases (α-amylase and amyloglucosidase) (α-G), inactive mutant laforin (C266S Laf) or wild type laforin (WT Laf) as indicated, and analyzed by SDS-Page (see Materials and Methods). Dried gels were analyzed by a Phosphorimager. C, control reaction lacking glycogen synthase; NT, not treated. B UDP-glucose and UDP were incubated with active (WT) or inactive (C266S) laforin as indicated and analyzed by HPAEC. Chromatograms of UDP, UMP and UDP-glucose standards are shown in the lowermost panel.

Article Snippet: High purity UDP (99.2%) used in laforin reactions was from Chem-Impex International, Inc. (#00310).

Techniques: Labeling, Incubation, Mutagenesis, SDS Page

Journal: Archives of biochemistry and biophysics

Article Title: INCORPORATION OF PHOSPHATE INTO GLYCOGEN BY GLYCOGEN SYNTHASE

doi: 10.1016/j.abb.2016.03.020

Figure Lengend Snippet: Release of 32Pi from 32P-glycogen by laforin. Glycogen was labeled by incubation with 5 μM [β-32P]UDP-glucose (A) or [U-14C]glucose (B) and yeast glycogen synthase (5 μg/ml) for 30 min. Glycogen was precipitated three times with ethanol, treated with PiBind™ resin, purified by gel filtration and dissolved in buffer. The glycogen was incubated for 2 hr with 50 μg/ml laforin (WT Laf) or laforin inactivated by boiling for 5 min (HI Laf). A control (C) lacked laforin. The reaction mixtures were analyzed by TLC using PEI-cellulose plates. Standards of glucose-1-P (14C-G1P), glucose (14C-Glu) and inorganic phosphate (32Pi), labeled with the indicated isotope, were also analyzed.

Article Snippet: High purity UDP (99.2%) used in laforin reactions was from Chem-Impex International, Inc. (#00310).

Techniques: Labeling, Incubation, Purification, Filtration