Determination of serum follicle stimulating hormone (FSH) level
Biotin-conjugated anti-FSH and standard or sample is incubated in
monoclonal anti-FSH antibody-coated wells. After 15~18
hours incubation and washing, HRP (horseradish peroxidase)-conjugated
avidin is added and incubated for 30 minutes. After washing, HRP-complex
remaining in wells are reacted with a chromogenic substrate (TMB) for 30
minutes, and reaction is stopped by addition of acidic solution, and
absorbance of yellow product is measured spectrophotometrically at 450
nm (sub-wavelength is 620nm). The absorbance is nearly proportional to
FSH concentration. The standard curve is prepared by plotting absorbance
against standard FSH concentrations
(Markkula, Hämäläinen, Loune, &
Huhtaniemi, 1995).