Human/Monkey Albumin Recombinant Rabbit Monoclonal Antibody [PSH20-16] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human Albumin matched pair antibodies

Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16]
Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724127) 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 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Human Albumin protein (A3782, sigma) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA724128, 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 SubstHumane 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.

Sandwich ELISA analysis of Monkey Albumin matched pair antibodies

Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16]
Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724127) diluted in carbonate/bicarbonate buffer, at a concentCynomolgusion 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 Monkey Albumin protein (BSA-CM101, Kactusbio) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA724128, 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 SubstHumane 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 human serum samples.

Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16]
Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17]

The concentrations of Albumin were measured in duplicates, interpolated from the Albumin standard curve and corrected for sample dilution. Undiluted samples are human serum 0.0000063%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Albumin concentration was determined to be 25.6 mg/ml in human serum.

Interpolated concentrations of native Albumin in HepG2 and HeLa cell culture supernatant.

Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16]
Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17]

The concentrations of Albumin were measured in duplicates, interpolated from the Albumin standard curve and corrected for sample dilution. Undiluted samples are HepG2 cell culture supernatant 0.5% and HeLa cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Albumin concentration was determined to be 745.1 ng/ml in HepG2 cell culture supernatant and undetectable in HeLa cell culture supernatant.