Biotin Conjugated Insulin Recombinant Rabbit Monoclonal Antibody [PSH04-16] - Detector

Sandwich ELISA analysis of Human Insulin matched pair antibodies

Capture: HA722096, Human Insulin Rabbit mAb [PSH04-15]
Detector: HA722097, Human Insulin Rabbit mAb [PSH04-16]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA722096) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant human Insulin (HA211183) protein starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA722097, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Interpolated concentrations of native Insulin in human serum and rat serum.

Capture: HA722096, Human Insulin Rabbit mAb [PSH04-15]
Detector: HA722097, Human Insulin Rabbit mAb [PSH04-16]

The concentrations of Insulin were measured in duplicates, interpolated from the Insulin standard curve and corrected for sample dilution. Undiluted samples are human serum 50% and rat serum 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Insulin concentration was determined to be 644 pg/ml in human serum and 614 pg/ml in rat serum.