Biotin Conjugated Human CSF-1-R Recombinant Rabbit Monoclonal Antibody [PSH12-66] - Detector

Sandwich ELISA analysis of Human CSF-1-R matched pair antibodies

Capture: HA723477, Human CSF-1-R Rabbit mAb [PSH12-68]
Detector: HA723474, Human CSF-1-R Rabbit mAb [PSH12-66]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723477) 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 CSF-1-R protein (HA211149) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723474, 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.

Interpolated concentrations of native CSF1R in THP-1 and Raji cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723477, Human CSF-1-R Rabbit mAb [PSH12-68]
Detector: HA723474, Human CSF-1-R Rabbit mAb [PSH12-66]

Interpolated concentration of native CSF1R was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CSF1R concentration was determined to be 3782 pg/mL in THP-1 cell extract and undetectable in Raji cell extract.

Interpolated concentrations of spiked CSF1R in cell culture media samples.

Capture: HA723477, Human CSF-1-R Rabbit mAb [PSH12-68]
Detector: HA723474, Human CSF-1-R Rabbit mAb [PSH12-66]

The concentrations of CSF1R were measured in duplicates, interpolated from the CSF1R 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).