Biotin Conjugated Human ANGPTL4 Recombinant Rabbit Monoclonal Antibody [PSH20-15] - Detector

Sandwich ELISA analysis of Human ANGPTL4 matched pair antibodies

Capture: HA725389, Human ANGPTL4 Rabbit mAb [PSH20-14]
Detector: HA725390, Human ANGPTL4 Rabbit mAb [PSH20-15]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725389) diluted in carbonate/bicarbonate buffer, at a concentration of 2 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 ANGPTL4 (HA211387) protein starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA725390, 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 ANGPTL4 in A549 cell culture supernatant and SW480 cell culture supernatant:

Capture: HA725389, Human ANGPTL4 Rabbit mAb [PSH20-14]
Detector: HA725390, Human ANGPTL4 Rabbit mAb [PSH20-15]

The concentrations of ANGPTL4 were measured in duplicates, interpolated from the ANGPTL4 standard curve and corrected for sample dilution. Undiluted samples are A549 cell culture supernatant 100% and SW480 cell culture supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ANGPTL4 concentration was determined to be 1472 pg/ml in A549 cell culture supernatant and 395 pg/ml in SW480 cell culture supernatant.