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Sandwich ELISA analysis of Mouse G-CSF matched pair antibodies
Capture: HA723777, Mouse G-CSF Rabbit mAb [PSH15-74]
Detector: HA723778, Mouse G-CSF Rabbit mAb [PSH15-75]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723777) 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 Mouse G-CSF protein (HA210972) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723778, 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 G-CSF in Raw264.7 LPS stimulated cell culture supernatant.
Capture: HA723777, Mouse G-CSF Rabbit mAb [PSH15-74]
Detector: HA723778, Mouse G-CSF Rabbit mAb [PSH15-75]
Raw264.7 cells were 24 hours serum starved and continued to culture either in the absence or in the presence of 5 µg/ml LPS for 48 hours. The concentrations of G-CSF measured in duplicate and interpolated from the G-CSF standard curve and corrected for sample dilution. Undiluted samples are as follows: unstimulated 100% and stimulated 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean G-CSF concentration was determined to be 1,277 pg/ml in LPS stimulated Raw264.7 cell culture supernatant and undetectable in the unstimulated Raw264.7 control.
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Interpolated concentrations of spiked G-CSF in mouse cell culture media samples.
Capture: HA723777, Mouse G-CSF Rabbit mAb [PSH15-74]
Detector: HA723778, Mouse G-CSF Rabbit mAb [PSH15-75]
The concentrations of G-CSF were measured in duplicates, interpolated from the G-CSF 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).
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