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Sandwich ELISA analysis of Mouse CD68 matched pair antibodies
Capture: HA723449, Mouse CD68 Rabbit mAb [PSH12-26]
Detector: HA723450, Mouse CD68 Rabbit mAb [PSH12-27]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723449) 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 CD68 protein (HA211131) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723450, 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 CD68 in mouse spleen, Raw264.7 and Neuro-2a extract samples based on a 1000 µg/ml extract load.
Capture: HA723449, Mouse CD68 Rabbit mAb [PSH12-26]
Detector: HA723450, Mouse CD68 Rabbit mAb [PSH12-27]
Interpolated concentration of native CD68 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD68 concentration was determined to be 35353 pg/mL in mouse spleen extract,47421 pg/ml in Raw264.7 extract and undetectable in Neuro-2a extract.
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Interpolated concentrations of spiked CD68 in cell culture media samples.
Capture: HA723449, Mouse CD68 Rabbit mAb [PSH12-26]
Detector: HA723450, Mouse CD68 Rabbit mAb [PSH12-27]
The concentrations of CD68 were measured in duplicates, interpolated from the CD68 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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