Biotin Conjugated Human Albumin Recombinant Rabbit Monoclonal Antibody [PSH27-50] - Detector

Sandwich ELISA analysis of Human Albumin protein matched pair antibodies

Capture: HA725335, Human Albumin Rabbit mAb [PSH27-49]
Detector: HA725336, Human Albumin Rabbit mAb [PSH27-50]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725335) 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 Albumin protein starting from 25,000 pg/mL to 0 pg/mL and detect antibody (HA725336, Biotin, 0.05 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 Albumin in HepG2 and HeLa cell supernatant.

Capture: HA725335, Human Albumin Rabbit mAb [PSH27-49]
Detector: HA725336, Human Albumin Rabbit mAb [PSH27-50]

The concentrations of native Albumin measured in duplicate and interpolated from the Albumin standard curve and corrected for sample dilution. Undiluted samples are HepG2 0.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Albumin concentration was determined to be 1,577.02 ng/mL in HepG2 cell culture supernatant. There was no detectable signal in HeLa cell culture supernatant.