Biotin Conjugated Mouse/Rat SCF Recombinant Rabbit Monoclonal Antibody [PSH16-92] - Detector

Sandwich ELISA analysis of Mouse SCF matched pair antibodies

Capture: HA723887, Mouse/Rat SCF Rabbit mAb [PSH16-91]
Detector: HA723888, Mouse/Rat SCF Rabbit mAb [PSH16-92]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723887) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 SCF protein (HA211067) starting from 1,000 pg/ml to 0 pg/ml and detect antibody (HA723888, 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.

Sandwich ELISA analysis of Mouse SCF matched pair antibodies

Capture: HA723887, Mouse/Rat SCF Rabbit mAb [PSH16-91]
Detector: HA723888, Mouse/Rat SCF Rabbit mAb [PSH16-92]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723887) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 SCF protein (HA211067) starting from 1,000 pg/ml to 0 pg/ml and detect antibody (HA723888, 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 SCF in mouse and rat samples.

Capture: HA723887, Mouse/Rat SCF Rabbit mAb [PSH16-91]
Detector: HA723888, Mouse/Rat SCF Rabbit mAb [PSH16-92]

Interpolated concentration of native SCF was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean SCF concentration was determined to be 1,179 pg/ml in rat serum，595 pg/mL in L929 cell supernatant and 64 pg/mL in J774A.1 cell supernatant.

Interpolated concentrations of native SCF in mouse samples.

Capture: HA723887, Mouse/Rat SCF Rabbit mAb [PSH16-91]
Detector: HA723888, Mouse/Rat SCF Rabbit mAb [PSH16-92]

Interpolated concentration of native SCF was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean SCF concentration was determined to be 4,551 pg/mL in L929 and 367 pg/mL in J774A.1 cell supernatant.

Interpolated concentrations of spiked SCF in cell culture media samples.

Capture: HA723887, Mouse/Rat SCF Rabbit mAb [PSH16-91]
Detector: HA723888, Mouse/Rat SCF Rabbit mAb [PSH16-92]

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

Interpolated concentrations of spiked SCF in cell culture media samples.

Capture: HA723887, Mouse/Rat SCF Rabbit mAb [PSH16-91]
Detector: HA723888, Mouse/Rat SCF Rabbit mAb [PSH16-92]

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