Biotin Conjugated Human FGL1 Recombinant Rabbit Monoclonal Antibody [PSH18-96] - Detector

Sandwich ELISA analysis of Human FGL1 matched pair antibodies

Capture: HA724032, Human FGL1 Rabbit mAb [PSH18-95]
Detector: HA724033, Human FGL1 Rabbit mAb [PSH18-96]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724032) 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 FGL1 protein starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724033, 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 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 FGL1 in HepG2 and Ramos cell culture supernatant.

Capture: HA724032, Human FGL1 Rabbit mAb [PSH18-95]
Detector: HA724033, Human FGL1 Rabbit mAb [PSH18-96]

Interpolated concentration of native FGL1 was measured in duplicate at different sample concentrations and interpolated from the FGL1 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FGL1 concentration was determined to be 165,366 pg/ml in HepG2 cell culture supernatant and undetectable in Ramos cell culture supernatant.

Interpolated concentrations of native FGL1 in HepG2 and Ramos Cell extract samples based on a 1000 µg/ml extract load.

Capture: HA724032, Human FGL1 Rabbit mAb [PSH18-95]
Detector: HA724033, Human FGL1 Rabbit mAb [PSH18-96]

Interpolated concentration of native FGL1 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FGL1 concentration was determined to be 26.309 pg/ml in HepG2 Cell extract and undetectable in Ramos Cell extract.

Interpolated concentrations of spiked FGL1 in cell culture media samples.

Capture: HA724032, Human FGL1 Rabbit mAb [PSH18-95]
Detector: HA724033, Human FGL1 Rabbit mAb [PSH18-96]

The concentrations of FGL1 were measured in duplicates, interpolated from the FGL1 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).