Human/Monkey VEGFR3 / FLT4 Recombinant Rabbit Monoclonal Antibody [PSH22-16] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human VEGFR3 matched pair antibodies

Capture: HA724277, Human/Monkey VEGFR3 / FLT4 Rabbit mAb [PSH22-16]
Detector: HA724278, Human/Monkey VEGFR3 / FLT4 Rabbit mAb [PSH22-17]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724277) diluted in carbonate/bicarbonate buffer, at a concentHumanion of 2 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.2% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human VEGFR3 / FLT4 protein (HA211107) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724278, Biotin, 0.2 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 SubstHumane for 10 minutes at room tempeHumanure in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Interpolated concentrations of native VEGFR3 in human serum samples and A549 cell culture supernatant.

Capture: HA724277, Human/Monkey VEGFR3 / FLT4 Rabbit mAb [PSH22-16]
Detector: HA724278, Human/Monkey VEGFR3 / FLT4 Rabbit mAb [PSH22-17]

The concentrations of VEGFR3 were measured in duplicates, interpolated from the VEGFR3 standard curve and corrected for sample dilution. Undiluted samples are human serum 5% and A549 cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGFR3 concentration was determined to be 138.3 ng/ml in human serum and undetectable in A549 cell culture supernatant.

Interpolated concentrations of spiked VEGFR3 in human cell culture media samples.

Capture: HA724277, Human/Monkey VEGFR3 / FLT4 Rabbit mAb [PSH22-16]
Detector: HA724278, Human/Monkey VEGFR3 / FLT4 Rabbit mAb [PSH22-17]

The concentrations of VEGFR3 were measured in duplicates, interpolated from the VEGFR3 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).