Biotin Conjugated Human Pentraxin 3 / PTX3 Recombinant Rabbit Monoclonal Antibody [PSH11-85] - Detector

Sandwich ELISA analysis of human Pentraxin 3 / PTX3 matched pair antibodies

Capture: HA723387, Human Pentraxin 3 / PTX3 Rabbit mAb [PSH11-83]
Detector: HA723390, Human Pentraxin 3 / PTX3 Rabbit mAb [PSH11-85]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723387) 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 Pentraxin 3 / PTX3 protein (HA210943) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723391B, 0.1 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 Pentraxin 3 in HUVEC cell culture supernatant.

Capture: HA723387, Human Pentraxin 3 / PTX3 Rabbit mAb [PSH11-83]
Detector: HA723390, Human Pentraxin 3 / PTX3 Rabbit mAb [PSH11-85]

The concentrations of Pentraxin 3 were interpolated from the Pentraxin 3 standard curve and corrected for sample dilution. Undiluted samples are HUVEC cell culture supernatant 100%. The mean Pentraxin 3 concentration was determined to be 308.1 pg/ml in HUVEC cell culture supernatant.

Interpolated concentrations of spiked Pentraxin 3 in human cell culture media samples.

Capture: HA723387, Human Pentraxin 3 / PTX3 Rabbit mAb [PSH11-83]
Detector: HA723390, Human Pentraxin 3 / PTX3 Rabbit mAb [PSH11-85]

The concentrations of Pentraxin 3 were measured in duplicates, interpolated from the Pentraxin 3 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).