Journal: The Journal of Biological Chemistry

Article Title: Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research *

doi: 10.1074/jbc.M112.396598

Figure Lengend Snippet: Microarray printing surfaces and reproducibility. Coupling oligosaccharides to protein enabled printing on a wide variety of slides and membranes commonly used for microarray production. (1→4)-β-d-Mannohexaose and (1→5)-α-l-arabinopentaose microarrays were printed in sextuplet and at 10 concentrations from 2 mg/ml to 30.5 ng/ml. The arrays were probed with the anti-mannan or anti-arabinan monoclonal antibodies LM21 and LM6, respectively. Microarrays were printed using a noncontact piezoelectric robot (A–F) or a pin-based robot (G and H). Arrays were printed on surface-modified glass slides (A–C), nitrocellulose-coated glass slides (D and E), and nitrocellulose membrane (F–H). Reproducibility of the microarrays was tested by printing 12 copies of arrays on both nitrocellulose membrane (I) and nitrocellulose-coated glass FAST Slides (J). Arrays were probed with the anti-mannan mAbs LM21 or the anti-arabinan mAb LM6. Representative probed replicate arrays are shown, as well as graphs of mean spot signals from three arrays plotted against each other. Axes on the graphs are relative mean spot signals.

Article Snippet: Carbohydrate microarrays were printed using two types of microarrays robot, a pin-based MicroGrid II (Digilab/Genomic Solutions, Huntingdon, UK) and a piezoelectric Sprint (Arrayjet, Roslin, UK).

Techniques: Microarray, Bioprocessing, Modification, Membrane

Journal: The Journal of Biological Chemistry

Article Title: Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research *

doi: 10.1074/jbc.M112.396598

Figure Lengend Snippet: Specificity screening of mAbs. Oligosaccharide microarrays were probed with a set of 38 monoclonal antibodies, and selected examples of probed arrays are shown in A. The mean spot signals obtained from three experiments are presented in heat maps in which color intensity is correlated to signal (B–I). Expanded heat maps (C–I) provide more detailed information about the binding of selected mAbs. The highest signal in the entire data set was set to 100, and all other values were normalized accordingly. Oligosaccharide structures are only shown where binding produced a mean spot signal of at least 15% of the highest mean signal in the entire data set. Further details about the oligosaccharides are provided in supplemental Table S1.

Article Snippet: Carbohydrate microarrays were printed using two types of microarrays robot, a pin-based MicroGrid II (Digilab/Genomic Solutions, Huntingdon, UK) and a piezoelectric Sprint (Arrayjet, Roslin, UK).

Techniques: Bioprocessing, Binding Assay, Produced

Journal: The Journal of Biological Chemistry

Article Title: Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research *

doi: 10.1074/jbc.M112.396598

Figure Lengend Snippet: Specificity screening of carbohydrate-binding modules and lectins. Oligosaccharide microarrays were probed with the xylan-binding CBM CBM4-2 and also mutant variants of CBM4-2: XG-34/1-X; XG-34/2-VI; X-6; X-13; M60-3; MRT-5; A-8; and A-12. The highest signal in the entire data set was set to 100, and all other values were normalized accordingly. Further details about the oligosaccharides are provided in supplemental Table S1.

Article Snippet: Carbohydrate microarrays were printed using two types of microarrays robot, a pin-based MicroGrid II (Digilab/Genomic Solutions, Huntingdon, UK) and a piezoelectric Sprint (Arrayjet, Roslin, UK).

Techniques: Binding Assay, Mutagenesis

Journal: The Journal of Biological Chemistry

Article Title: Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research *

doi: 10.1074/jbc.M112.396598

Figure Lengend Snippet: Oligosaccharide microarrays as multiplexed acceptors for enzyme assays. A, schematic showing the experimental setup used to assay the activity of rabbit muscle phosphorylase using a [14C]glucose 1-phosphate donor and an array of immobilized oligosaccharide acceptor. B, an oligosaccharide microarray was used to assay the activity of rabbit muscle phosphorylase a with [14C]glucose 1-phosphate as a glycosyl donor. The incorporation of 14C was detected using a PhosphorImager system. The control array in C was probed with boiled phosphorylase a under the same conditions. The structures of the oligosaccharides that were the most effective acceptors are shown. Further details about the oligosaccharides are provided in supplemental Table S1.

Article Snippet: Carbohydrate microarrays were printed using two types of microarrays robot, a pin-based MicroGrid II (Digilab/Genomic Solutions, Huntingdon, UK) and a piezoelectric Sprint (Arrayjet, Roslin, UK).

Techniques: Activity Assay, Microarray, Control