Biotin Conjugated Human GCSF Receptor Recombinant Rabbit Monoclonal Antibody [PSH18-04] - Detector

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

Capture: HA725291, Human GCSF Receptor Rabbit mAb [PSH18-03]
Detector: HA725292, Human GCSF Receptor Rabbit mAb [PSH18-04]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725291) 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 GCSF Receptor protein (HA211262) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA725292, 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 G-CSF-R in THP-1 and MCF7 extract samples based on a 1,000 µg/ml extract load.

Capture: HA725291, Human GCSF Receptor Rabbit mAb [PSH18-03]
Detector: HA725292, Human GCSF Receptor Rabbit mAb [PSH18-04]

The concentrations of G-CSF-R were measured in duplicates, interpolated from the G-CSF-R standard curve and corrected for sample dilution. Undiluted samples are THP-1 extract 100% and MCF7 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean G-CSF-R concentration was determined to be 525.2 pg/ml in THP-1 extract and undetectable in MCF7 extract.