Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Protein-specific analysis of invertebrate glycoproteins

doi: 10.1007/978-1-4939-8814-3_24

Figure Lengend Snippet: List of selected antibodies, lectins and pentraxins for N-glycan epitope screening ( Note 4 )

Article Snippet: SDS-PAGE and Western blotting list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 12% SDS-PAGE gel (using 40% acrylamide stock from Bio-Rad, diluted with either stacking gel buffer with 0.5 M Tris/HCl pH 6.8 or separation gel buffer with 1.5 M Tris/HCl pH 8.8) SDS-PAGE running buffer (25 mM Tris, 192 mM glycine, 0.1% SDS; components from either VWR or Roth) Western blotting transfer buffer (25 mM Tris, 192 mM glycine, 10% methanol; VWR or Roth) SDS-PAGE protein standard ladder (e.g., Thermo PageRuler™) Nitrocellulose membrane (BioTrace™ NT from Pall Life Science) Extra thick blotting paper (Bio-Rad) 0.5 % (w/v) Ponceau S (Sigma) in 1 % (v/v) acetic acid solution Membrane washing buffer: Tris buffered saline (TBS, i.e., 0.1 M Tris/HCl, pH 7.4, 0.1 M NaCl; typically made as a 10-fold concentrated stock) with 0.05% Tween (Sigma) Membrane blocking and antibody/lectin dilution buffer: Tris buffered saline with 0.05% Tween and 0.5% BSA (Roth) Primary, secondary antibodies, lectins and pentraxins (Sigma or Vector Laboratories) (see ) table ft1 table-wrap mode="anchored" t5 caption a7 Antibody (1 st ) Dilution Epitope( 13 , 14 ) Supplier Anti-HRP from rabbit, 10 mg/ml 1:10000 Core α1,3-Fuc/ core β1,2-Xyl Sigma Anti-PC (TEPC-15 mouse IgA), 10 mg/ml 1:200 PC-Hex(NAc) Sigma Antibody (2 nd ) Anti-rabbit IgG from goat conjugated with alkaline phosphatase 1:2000 Vector Labs Anti-mouse IgA from goat conjugated with alkaline phosphatase 1:10000 Sigma Pentraxin C reactive Protein (CRP) from human plasma (CaCl 2 2.5 mM added) 1:200 PC-Hex(NAc) MP Biochemicals Amyloid P component from human serum (SAP) 1:200 PE-Hex(NAc) Sigma Pentraxin recognition Anti-human C reactive protein from rabbit 1:1000 Dako Anti-Amyloid P human serum component IgG from rabbit (anti SAP) 1:1000 Calbiochem Lectin Biotinylated Aleuria aurantia lectin 1:1000 Core α1,6-Fuc/Lex Vector Labs Biotinylated wheat germ agglutinin 1:1000 β1,4HexNAc/α2,3Sia Vector Labs Biotinylated peanut agglutinin 1:1000 Galβ1,3GalNAc Vector Labs Lectin recognition Anti-biotin from goat conjugated with alkaline phosphatase 1:10000 Sigma Open in a separate window caption a8 List of selected antibodies, lectins and pentraxins for N-glycan epitope screening ( Note 4 ) SigmaFAST BCIP/NBT or SigmaFAST 3,3´-diaminobenzidine tetrahydrochloride tablets (Sigma), dissolved in 10ml and 5ml respectively.

Techniques: Plasmid Preparation

Journal: Glycoconjugate Journal

Article Title: Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients

doi: 10.1007/s10719-013-9501-y

Figure Lengend Snippet: Relative reactivity of seminal plasma IgG with fucose-specific lectins

Article Snippet: Three biotinylated fucose-specific lectins (Vector Laboratories Inc., Burlingame, CA, USA): Aleuria aurantia lectin (AAL), Ulex europaeus agglutinin (UEA) and Lotus tetragonolobus agglutinin (LTA) were used to determine expression of fucose moiety in IgG by lectin-ELISA according to the procedure described previously for fibronectin and α 1 -acid glycoprotein [ ].

Techniques:

Journal: Glycoconjugate Journal

Article Title: Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients

doi: 10.1007/s10719-013-9501-y

Figure Lengend Snippet: IgG relative reactivity with fucose-specific lectins. The relative reactivity of seminal IgG with biotinylated fucose-specific lectins AAL, UEA and LTA was determined in normal and leukocytospermic groups using lectin-ELISA and expressed in absorbance units (AU). □ – median value

Article Snippet: Three biotinylated fucose-specific lectins (Vector Laboratories Inc., Burlingame, CA, USA): Aleuria aurantia lectin (AAL), Ulex europaeus agglutinin (UEA) and Lotus tetragonolobus agglutinin (LTA) were used to determine expression of fucose moiety in IgG by lectin-ELISA according to the procedure described previously for fibronectin and α 1 -acid glycoprotein [ ].

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Glycoconjugate Journal

Article Title: Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients

doi: 10.1007/s10719-013-9501-y

Figure Lengend Snippet: Dependence between IgG concentration and IgG relative reactivity with lectins. Correlation between IgG concentration and IgG relative reactivity with AAL ( a ), and UEA ( b ) was estimated according to a Spearman test, with a two-tailed p -value of less than 0.05 considered significant. 95 % confidence interval is marked by dotted lines

Article Snippet: Three biotinylated fucose-specific lectins (Vector Laboratories Inc., Burlingame, CA, USA): Aleuria aurantia lectin (AAL), Ulex europaeus agglutinin (UEA) and Lotus tetragonolobus agglutinin (LTA) were used to determine expression of fucose moiety in IgG by lectin-ELISA according to the procedure described previously for fibronectin and α 1 -acid glycoprotein [ ].

Techniques: Concentration Assay, Two Tailed Test

Journal: Glycoconjugate Journal

Article Title: Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients

doi: 10.1007/s10719-013-9501-y

Figure Lengend Snippet: Relative reactivity of SC with fucose-specific lectins. Lectin-blotting with three biotinylated fucose-specific lectins AAL, UEA and LTA was performed for normal ( 1 ) and leukocytospermic ( 2 ) seminal groups. The bands corresponding to SC (78 ± 4.2 kDa and 63 ± 2.3 kDa) are shown by arrows

Article Snippet: Three biotinylated fucose-specific lectins (Vector Laboratories Inc., Burlingame, CA, USA): Aleuria aurantia lectin (AAL), Ulex europaeus agglutinin (UEA) and Lotus tetragonolobus agglutinin (LTA) were used to determine expression of fucose moiety in IgG by lectin-ELISA according to the procedure described previously for fibronectin and α 1 -acid glycoprotein [ ].

Techniques:

Journal: Glycoconjugate Journal

Article Title: Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients

doi: 10.1007/s10719-013-9501-y

Figure Lengend Snippet: IgG relative reactivity with fucose-specific lectins. The relative reactivity of seminal IgG with biotinylated fucose-specific lectins AAL, UEA and LTA was determined in normal and leukocytospermic groups using lectin-ELISA and expressed in absorbance units (AU). □ – median value

Article Snippet: Three biotinylated fucose-specific lectins (Vector Laboratories Inc., Burlingame, CA, USA): Aleuria aurantia lectin (AAL), Ulex europaeus agglutinin (UEA) and Lotus tetragonolobus agglutinin (LTA) were used to determine expression of fucose moiety in IgG by lectin-ELISA according to the procedure described previously for fibronectin and α 1 -acid glycoprotein [ ].

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Glycoconjugate Journal

Article Title: Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients

doi: 10.1007/s10719-013-9501-y

Figure Lengend Snippet: Relative reactivity of SC with fucose-specific lectins. Lectin-blotting with three biotinylated fucose-specific lectins AAL, UEA and LTA was performed for normal ( 1 ) and leukocytospermic ( 2 ) seminal groups. The bands corresponding to SC (78 ± 4.2 kDa and 63 ± 2.3 kDa) are shown by arrows

Article Snippet: Three biotinylated fucose-specific lectins (Vector Laboratories Inc., Burlingame, CA, USA): Aleuria aurantia lectin (AAL), Ulex europaeus agglutinin (UEA) and Lotus tetragonolobus agglutinin (LTA) were used to determine expression of fucose moiety in IgG by lectin-ELISA according to the procedure described previously for fibronectin and α 1 -acid glycoprotein [ ].

Techniques: