Human TNF Receptor II Recombinant Rabbit Monoclonal Antibody [PSH11-94] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human TNF-R2 matched pair antibodies

Capture: HA723405, Human TNF Receptor II Rabbit mAb [PSH11-94]
Detector: HA723403, Human TNF Receptor II Rabbit mAb [PSH11-93]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723405) 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 TNF Receptor II protein (HA210950) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723403, 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 TNF-R2 in U937, SW620 and HeLa cell culture supernatant.

Capture: HA723405, Human TNF Receptor II Rabbit mAb [PSH11-94]
Detector: HA723403, Human TNF Receptor II Rabbit mAb [PSH11-93]

The concentrations of TNF-R2 were measured in duplicates, interpolated from the TNF-R2 standard curve and corrected for sample dilution. Undiluted samples are U937 cell culture supernatant 100%, SW620 cell culture supernatant 100% and HeLa cell culture supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF-R2 concentration was determined to be 192.5 pg/ml in U937 cell culture supernatant, 569.2 pg/ml in SW620 cell culture supernatant and undetectable in HeLa cell culture supernatant.

Interpolated concentrations of native TNF-R2 in human urine samples.

Capture: HA723405, Human TNF Receptor II Rabbit mAb [PSH11-94]
Detector: HA723403, Human TNF Receptor II Rabbit mAb [PSH11-93]

The concentrations of TNF-R2 were measured in duplicates, interpolated from the TNF-R2 standard curve and corrected for sample dilution. Undiluted samples are human urine 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF-R2 concentration was determined to be 8,928 pg/ml in human urine.

Interpolated concentrations of spiked TNF-R2 in human cell culture media samples.

Capture: HA723405, Human TNF Receptor II Rabbit mAb [PSH11-94]
Detector: HA723403, Human TNF Receptor II Rabbit mAb [PSH11-93]

The concentrations of TNF-R2 were measured in duplicates, interpolated from the TNF-R2 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).