Biotin Conjugated Human/Monkey IGFBP2 Recombinant Rabbit Monoclonal Antibody [PSH25-34] - Detector

Sandwich ELISA analysis of Human IGFBP2 matched pair antibodies

Capture: HA724524, Human/Monkey IGFBP2 Rabbit mAb [PSH25-33]
Detector: HA724525, Human/Monkey IGFBP2 Rabbit mAb [PSH25-34]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724524) 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 IGFBP2 protein (HA211103) starting from 40,000 pg/mL to 0 pg/mL and detect antibody (HA724525, Biotin, 0.05 μg/mL) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 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 IGFBP2 in human and monkey samples.

Capture: HA724524, Human/Monkey IGFBP2 Rabbit mAb [PSH25-33]
Detector: HA724525, Human/Monkey IGFBP2 Rabbit mAb [PSH25-34]

The concentrations of IGFBP2 were measured in duplicates, interpolated from the IGFBP2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IGFBP2 concentration was determined to be 554,351 pg/mL in Cynomolgus serum, 213,887 pg/mL in human serum and 55,588 pg/mL in T-47D cell supernatant.

Interpolated concentrations of spiked IGFBP2 in human cell culture media samples.

Capture: HA724524, Human/Monkey IGFBP2 Rabbit mAb [PSH25-33]
Detector: HA724525, Human/Monkey IGFBP2 Rabbit mAb [PSH25-34]

The concentrations of IGFBP2 were measured in duplicates, interpolated from the IGFBP2 standard curves and corrected for sample dilution. Diluted samples are as follows: 25% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).