Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: 'ZP domain' of human zona pellucida glycoprotein-1 binds to human spermatozoa and induces acrosomal exocytosis

doi: 10.1186/1477-7827-8-110

Figure Lengend Snippet: Profile of lectins binding to baculovirus-expressed human ZP1 273-551aa in ELISA . Microtitration plates were coated with the baculovirus-expressed recombinant ZP1 273-551aa (open bar; 500 ng/well) and processed for evaluation of binding to 21 different biotinylated lectins in an ELISA as described in Methods . As an internal control, microtitration plates coated with same amount of E. coli -expressed recombinant human ZP1 273-551aa (grey bar; unpublished observations) were also used. Values are expressed as absorbance obtained with various lectins binding to the respective recombinant protein, after deducting the non-specific binding of the lectins to the uncoated wells. Each bar represents a mean of duplicate experiments and standard deviation of the absorbance values. The lectins tested were GSL-I: Griffon simplicifolia lectin I, SBA: Soybean agglutinin, DSL: Datura stramonium lectin, PSA: Pisum sativum agglutinin, ConA: Concanavalin A, ECL: Erythrina cristagalli lectin, LCA: Lens culinaris agglutinin, DBA: Dolichos biflorus agglutinin, LEL: Lycopersicon esculentum lectin, PHA-L: Phaseolus vulgaris leucoagglutinin, UEA-I: Ulex europaeus agglutinin I, STL: Solanum tuberosum lectin, PHA-E: Phaseolus vulgaris erythroagglutinin, RCA: Ricinus communis agglutinin, VVA: Vicia villosa agglutinin, SJA: Sophora japonica agglutinin, PNA: Peanut agglutinin, S- WGA: Succinylated Wheat germ agglutinin, Jacalin, WGA: Wheat Germ agglutinin and GSL II: Griffonia simplicifolia lectin II.

Article Snippet: The plate was blocked with 0.1% Tween-20 in PBS (PBST, 200 μl/well) for 90 min at 37°C followed by incubation with 21 biotinylated lectins (1 μg/ml; 100 μl/well) at 37°C for 1 h. The biotinylated lectins available in the Lectin kit-I, II and III (Vector Laboratories, Burlingame, CA, USA) were used in the lectin binding assay.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation