EPO Recombinant Rabbit Monoclonal Antibody [PSH24-53] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human EPO matched pair antibodies

Capture: HA725351, EPO Rabbit mAb [PSH24-52]
Detector: HA725352, EPO Rabbit mAb [PSH24-53]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725351) diluted in carbonate/bicarbonate buffer, at a concentration of 3 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 EPO protein (HA210852) starting from 500 pg/ml to 0 pg/ml and detect antibody (HA725352, 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.

Sandwich ELISA analysis of Mouse EPO matched pair antibodies

Capture: HA725351, EPO Rabbit mAb [PSH24-52]
Detector: HA725352, EPO Rabbit mAb [PSH24-53]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725351) diluted in carbonate/bicarbonate buffer, at a concentration of 3 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 Mouse EPO protein (HA211068) starting from 500 pg/ml to 0 pg/ml and detect antibody (HA725352, 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 EPO in human plasma and HeLa supernatant.

Capture: HA725351, EPO Rabbit mAb [PSH24-52]
Detector: HA725352, EPO Rabbit mAb [PSH24-53]

The concentrations of EPO were measured in duplicates, interpolated from the EPO standard curve and corrected for sample dilution. Undiluted samples are 50% human plasma and 50% HeLa supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean EPO concentration was determined to be 57.2 pg/ml in human plasma.

Interpolated concentrations of native EPO in mouse plasma and rat plasma.

Capture: HA725351, EPO Rabbit mAb [PSH24-52]
Detector: HA725352, EPO Rabbit mAb [PSH24-53]

The concentrations of EPO were measured in duplicates, interpolated from the EPO standard curve and corrected for sample dilution. Undiluted samples are 50% mouse plasma and 50% rat supernatant . The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean EPO concentration was determined to be 97 pg/ml in mouse plasma, 83 pg/ml in rat plasma.