Biotin Conjugated Human MERTK Recombinant Rabbit Monoclonal Antibody [PSH12-70] - Detector

Sandwich ELISA analysis of Human MERTK matched pair antibodies

Capture: HA723479, Human MERTK Rabbit mAb [PSH12-69]
Detector: HA723480, Human MERTK Rabbit mAb [PSH12-70]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723479) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 MERTK protein (HA211172) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723480, 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 MERTK in HepG2 and MCF7 extract samples based on a 1000 µg/ml extract load.

Capture: HA723479, Human MERTK Rabbit mAb [PSH12-69]
Detector: HA723480, Human MERTK Rabbit mAb [PSH12-70]

Interpolated concentration of native MERTK was measured in duplicate at different sample concentrations and interpolated from the MERTK standard curves. The mean MERTK concentration was determined to be 1,799 pg/mL in Hep G2 cell extract, There was no detectable signal in MCF7 cell extract.

Interpolated concentrations of native MERTK in human samples.

Capture: HA723479, Human MERTK Rabbit mAb [PSH12-69]
Detector: HA723480, Human MERTK Rabbit mAb [PSH12-70]

Interpolated concentration of native MERTK was measured in duplicate at different sample concentrations. Undiluted samples were 10% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean MERTK concentration was determined to be 119,371 pg/mL in human serum.

Interpolated concentrations of spiked MERTK in cell culture media samples.

Capture: HA723479, Human MERTK Rabbit mAb [PSH12-69]
Detector: HA723480, Human MERTK Rabbit mAb [PSH12-70]

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