Journal: Molecular & Cellular Proteomics : MCP

Article Title: Deciphering Protein Glycosylation by Computational Integration of On-chip Profiling, Glycan-array Data, and Mass Spectrometry *

doi: 10.1074/mcp.RA118.000906

Figure Lengend Snippet: Biological reagents

Article Snippet: Datura stramonium lectin (DSL) , Lectin , Vector Laboratories , BK-3000.

Techniques: Plasmid Preparation, Purification

Journal: Proteomics

Article Title: Identification of candidate biomarkers with cancer-specific glycosylation in the tissue and serum of endometrioid ovarian cancer patients by glycoproteomic analysis

doi: 10.1002/pmic.200900537

Figure Lengend Snippet: Tissue validation. (A) Lectin blot analysis of POSTN immunoprecipitations. POSTN was immunoprecitated from 500 μg of total cell lysate using a polyclonal antibody (Abcam, Cambridge, MA) prior to separation on 4–12% polyacrylamide gel and transfer to PVDF membrane. Blots were probed with biotinylated lectins (1:5000) and detected using streptavidin-coupled HRP (1:5000) and chemiluminescent development. (B) Lectin blot analysis of LAMP-1 immunoprecipitation reactions. LAMP-1 was immunoprecipitated from 500 μg of total cell lysates (normal-NL and tumor-TU) using a monoclonal antibody (E-Biosciences, San Diego, CA) detected by lectin blot as described above.

Article Snippet: The sample was adjusted to 150 mM NaCl, 5 mM CaCl 2 , and 5 mM MgCl 2 before the addition of the following lectins: biotinylated E-PHA ( Phaseolus vulgaris erythroagglutinin ), biotinylated AAL ( Aleuria aurantia ), and biotinylated DSL ( Datura stramonium ) (Vector Labs, Burlingame, CA) (10 μg each) were added and the sample was rotated at 4°C overnight.

Techniques: Immunoprecipitation

Journal: The Journal of biological chemistry

Article Title: The glycosylation of the influenza A virus hemagglutinin by mammalian cells. A site-specific study.

doi: 10.1074/jbc.272.7.4027

Figure Lengend Snippet: FIG. 4. Serial lectin affinity chromatography of neutral oligo- saccharides from individual glycosylation sites containing com- plex-type glycans. Elution profiles are from A. aurantia lectin (AAL) (a, d, and g), with subsequent fractionation of the major fractions from A. aurantia lectin on D. stramonium lectin (DSA) (b, e, and h), and from D. stramonium lectin on G. simplicifolia lectin I (GSL-I) (c, f, and i). Representative elution profiles are shown for each site. Site 129 is the Fo variant (a–c); site 21 is the Cf variant (d–f); and site 271 is the Co variant (g–i). 2, unbound material; 2/1, retarded material; 1, indi- cates strongly bound material. Arrows indicate the start of elution of strongly bound glycans with fucose (A. aurantia lectin), GlcNAc oli- gomers (D. stramonium lectin), or a-methylgalactoside ( G. simplicifolia lectin I).

Article Snippet: Immobilized Lectin Affinity Chromatography—Agarose-bound Aleuria aurantia lectin and Datura stramonium lectin, both from Vector Laboratories (Peterborough, UK) were packed into 5 3 50-mm glass columns (Pharmacia).

Techniques: Affinity Chromatography, Glycoproteomics, Fractionation, Variant Assay