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Sandwich ELISA analysis of Mouse M-CSF/CSF1 matched pair antibodies
Capture: HA723792, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-84]
Detector: HA723793, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-85]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723792) 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 Mouse M-CSF/CSF1 protein (HA211182) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723793, 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 M-CSF in mouse LPS and recombinant mouse ING-γ unstimulated and stimulated RAW264.7 cell culture supernatant.
Capture: HA723792, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-84]
Detector: HA723793, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-85]
RAW264.7 cells were stimulated with 1 µg/ml LPS and 30 ng/ml recombinant mouse ING-γ or vehicle control and incubated for 24 hours. The concentrations of M-CSF measured in duplicate and interpolated from the M-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 M-CSF concentration was determined to be 731.4 pg/ml in LPS and recombinant mouse ING-γ stimulated RAW264.7 cell culture supernatant and undetectable in the unstimulated RAW264.7 control.
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Interpolated concentrations of native M-CSF in mouse LPS and recombinant mouse ING-γ unstimulated and stimulated J774A.1 cell culture supernatant.
Capture: HA723792, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-84]
Detector: HA723793, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-85]
J774A.1 cells were stimulated with 1 µg/ml LPS and 100 ng/ml recombinant mouse ING-γ or vehicle control and incubated for 72 hours. The concentrations of M-CSF measured in duplicate and interpolated from the M-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 M-CSF concentration was determined to be 704.2 pg/ml in LPS and recombinant mouse ING-γ stimulated J774A.1 cell culture supernatant and undetectable in the unstimulated J774A.1 control.
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Interpolated concentrations of spiked M-CSF in mouse cell culture media samples.
Capture: HA723792, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-84]
Detector: HA723793, Mouse M-CSF/CSF1 Rabbit mAb [PSH15-85]
The concentrations of M-CSF were measured in triplicates, interpolated from the M-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=3).
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