The invention belongs to the technical field of biological
medicine, and particularly relates to a
kidney injury biomarker
protein detection method which comprises the following specific steps: S1, acquiring a
urine sample to be detected, and centrifuging to obtain supernate; s2, adding 1 [mu] g / ml of PBS of a goat anti-
rabbit antibody into an ELISA reaction plate, and then carrying out sealed incubation on the ELISA reaction plate;
drying the reaction plate; washing with PBS (
Phosphate Buffer Solution) (PBST) containing 0.2% of Twain-20, beating the reaction plate to dry, and repeating for two times to obtain a coated ELISA (
Enzyme-Linked Immunosorbent
Assay) plate; s3, respectively dissolving DKK3 and beta2-MG
protein standard substances with PBS, respectively diluting, and drawing standard curves for the
protein standard solutions with different concentrations; s4, respectively and sequentially adding 50 microliters of protein standard solutions or detection
urine samples with different concentrations, 100 microliters of HRP labeled antibodies, 100 microliters of TMB developing solutions and 50 microliters of stop solutions into the coated ELISA plate holes; s5, within 15 minutes after the reaction is terminated, putting the reaction plate into a
microplate reader, and reading an OD450 value; and S6, drawing a
standard curve, and calculating the concentrations of DKK3 and beta2-MG proteins according to the
standard curve.