A Highly Sensitive Sandwich Enzyme Immunoassay for Insulin in Human Serum Developed Using Capybara Anti-Insulin Fab' -Horseradish Peroxidase Conjugate

A noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for salmon calcitonin: Determination of the bioavailability of subcutaneous salmon calcitonin and its correlation with the hypocalcemic activity in rats

“…2) Nonspecifically bound peroxidase activity (4.0 f 0.40 SD) was measured using the 10 plasma samples along with SCT standard, which had been dissolved in the pooled plasma (Table 1, left column). an antigen to the antibody-coated solid phase, and/or the binding of an enzyme-labeled antibody to the antigen, which is trapped onto the solid phase (13,17). Serum (plasma) interference can be eliminated with increasing concentrations of inorganic salts (NaC1, KCl, 0.4 mol/l is sufficient) during incubation of serum (plasma) samples with the solid phase (1 3, 17).…”

“…After washing as described above. the peroxidase activity bound to the balls was assayed by fluorometry (13). The balls were preincubated with 6.0 g/l3-(4-hydroxypheny1)propionic acid (Tokyo Kasei Kogyo, Co., Ltd., Tokyo, Japan) in 0.1 mol/l sodium phosphate buffer, pH 7.0 (0.1 ml), at 30°C for 5 minutes.…”

Development of a highly sensitive and specific two-site enzyme immunoassay for parathyroid hormone (1-34): Application to pharmacokinetic study on intranasal parathyroid hormone (1-34) in human

“…The scales of 0 and 100 were adjusted by using 0.1 mol/l glycine-NaOH buffer, pH 10.3 and 10 -8 mol/l 4-methylumbelliferone (Nacalai Tesque, Inc.) in the same buffer, respectively. The fluorescence intensity was measured relative to the 4-methylumbelliferone solution (25).…”

Determination of the bioavailability of intranasal elcatonin in humans: Development of a sandwich transfer enzyme immunoassay for elcatonin

“…The scales of 0 and 100 were adjusted by using 0.1 mol/l glycine-NaOH buffer, pH 10.3, and 10 -8 mol/l 4-methylumbelliferone (Nakalai Tesque, Inc.) in the same buffer, respectively. The fluorescence intensity was measured relative to the 4-methylumbelliferone solution (20).…”

“…The colored polystyrene balls were removed, and the incubation was continued at 20°C for 2 hr. The white polystyrene balls were washed as above, and β-D-galactosidase activity bound to the white polystyrene balls was assayed by fluorometry (20) as follows. The white balls were preincubated with 10 mmol/l sodium phosphate buffer, pH 7.0, containing 0.1 mol/l NaCl, 1 mmol/l MgCl 2 , 1 g/l NaN 3 , and 0.1 g/l bovine serum albumin (0.1 ml) at 30°C for 5 min.…”