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Sandwich ELISA analysis of Human PARK7 matched pair antibodies
Capture: HA620001 Human PARK7 Mouse mAb [PSH15-67]
Detector: HA620002, Human PARK7 Mouse mAb [PSH15-68]
Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA620001) 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 PARK7 protein (HA211158) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA620002, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 PARK7 in 0.125 mg/ml HL-60 lysate samples.
Capture: HA620001 Human PARK7 Mouse mAb [PSH15-67]
Detector: HA620002, Human PARK7 Mouse mAb [PSH15-68]
The concentrations of PARK7 were measured in duplicates, interpolated from the PARK7 standard curve and corrected for sample dilution. Undiluted samples are 0.125 mg/ml HL-60 lysate . The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PARK7 concentration was determined to be 64.8 ng/ml in HL-60 lysate.
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