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Sandwich ELISA analysis of Human PDGFR alpha matched pair antibodies
Capture: HA725515, Human PDGFR alpha Rabbit mAb [PSH26-33]
Detector: HA725516, Human PDGFR alpha Rabbit mAb [PSH26-34]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725515) 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 PDGFR alpha protein (HA211519) starting from 5,000 pg/mL to 0 pg/mL and detect antibody (HA725516, 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.
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Interpolated concentrations of native PDGFR alpha in human serum samples.
Capture: HA725515, Human PDGFR alpha Rabbit mAb [PSH26-33]
Detector: HA725516, Human PDGFR alpha Rabbit mAb [PSH26-34]
The concentrations of PDGFR alpha were measured in duplicates, interpolated from the PDGFR alpha standard curve and corrected for sample dilution. Undiluted samples are human serum 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PDGFR alpha concentration was determined to be 5,414 pg/mL in human serum.
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