Biotin Conjugated Human c-Kit Recombinant Rabbit Monoclonal Antibody [PSH26-42] - Detector

Sandwich ELISA analysis of Human c-Kit matched pair antibodies

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724619) 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 Human c-Kit protein (HA211233) starting from 3,000 pg/mL to 0 pg/mL and detect antibody (HA724621, 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 c-Kit in IMR-32 cell extract samples based on a 1,000 µg/mL extract load.

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42]

Interpolated concentration of native c-Kit was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Kit concentration was determined to be 7,935 pg/mL in IMR-32 cell extract.

Interpolated concentrations of native c-Kit in human serum.

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42]

Interpolated concentration of native c-Kit was measured in duplicate at different sample concentrations and interpolated from the c-Kit standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Kit concentration was determined to be 163,987 pg/mL in human serum.

Interpolated concentrations of spiked c-Kit in cell culture media samples.

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42]

The concentrations of c-Kit were measured in duplicates, interpolated from the c-Kit 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=2).