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Sandwich ELISA analysis of Human GLP-1 matched pair antibodies
Capture: HA725353, Human/Mouse GLP-1 Rabbit mAb [PSH23-57]
Detector: HA725354, Human/Mouse GLP-1 Rabbit mAb [PSH23-58]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725353) 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 GLP-1 protein starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725354, Biotin, 0.5 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 GLP-1 in human serum samples.
Capture: HA725353, Human/Mouse GLP-1 Rabbit mAb [PSH23-57]
Detector: HA725354, Human/Mouse GLP-1 Rabbit mAb [PSH23-58]
Four individual healthy donors were evaluated for the presence of GLP-1 in serum using this assay. Serum from each donor was used undiluted without prior dilution. The mean levels of GLP-1 were found to be 43.9 pg/mL.
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Interpolated concentrations of native GLP-1 in mouse pancreas extract.
Capture: HA725353, Human/Mouse GLP-1 Rabbit mAb [PSH23-57]
Detector: HA725354, Human/Mouse GLP-1 Rabbit mAb [PSH23-58]
The concentrations of GLP-1 were measured in duplicates, interpolated from the GLP-1 standard curve and corrected for sample dilution. Undiluted samples are mouse pancreas extract 50 μg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean GLP-1 concentration was determined to be 1,888 pg/ml in mouse pancreas extract.
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