Human Heme Oxygenase 1 (HO-1) Recombinant Rabbit Monoclonal Antibody [PSH27-90] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human Heme Oxygenase 1 (HO-1) protein matched pair antibodies

Capture: HA724732, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-90]
Detector: HA724733, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-91]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724732) 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 Heme Oxygenase 1 (HO-1) protein (HA211372) starting from 2,500 pg/mL to 0 pg/mL and detect antibody (HA724733, 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 Heme Oxygenase 1 (HO-1) in A549 and HeLa extract samples based on a 1,000 µg/mL extract load.

Capture: HA724732, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-90]
Detector: HA724733, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-91]

Interpolated concentration of native Heme oxygenase 1 (HO-1) was measured in duplicate at different sample concentrations and interpolated from the Heme oxygenase 1 (HO-1) standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Heme oxygenase 1 (HO-1) concentration was determined to be 24,004 pg/mL in A549 cell extract and 7,189 pg/mL in HeLa cell extract.

Interpolated concentrations of native Heme Oxygenase 1 (HO-1) in human samples.

Capture: HA724732, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-90]
Detector: HA724733, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-91]

Interpolated concentration of native Heme Oxygenase 1 (HO-1) was measured in duplicate at different sample concentrations and interpolated from the Heme Oxygenase 1 (HO-1) standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Heme Oxygenase 1 (HO-1) concentration was determined to be 551 pg/mL in human serum.

Interpolated concentrations of spiked Heme Oxygenase 1 (HO-1) in cell culture media samples.

Capture: HA724732, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-90]
Detector: HA724733, Human Heme Oxygenase 1 (HO-1) Rabbit mAb [PSH27-91]

The concentrations of Heme Oxygenase 1 (HO-1) were measured in duplicates, interpolated from the Heme Oxygenase 1 (HO-1) 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).