Biotin Conjugated Human/Mouse c-Fos Recombinant Rabbit Monoclonal Antibody [PSH17-93] - Detector

Sandwich ELISA analysis of Human/Mouse c-Fos matched pair antibodies

Capture: HA723957, Human/Mouse c-Fos Rabbit mAb [PSH17-92]
Detector: HA723958, Human/Mouse c-Fos Rabbit mAb [PSH17-93]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723957) 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 Human c-Fos protein (HA211326) starting from 3000 pg/ml to 0 pg/ml and detect antibody (HA723958, 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-Fos in HeLa treated or untreated with starve 16 hours then TPA for 4 hours cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723957, Human/Mouse c-Fos Rabbit mAb [PSH17-92]
Detector: HA723958, Human/Mouse c-Fos Rabbit mAb [PSH17-93]

Interpolated concentration of native c-Fos was measured in duplicate at different sample concentrations and interpolated from the c-Fos standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Fos concentration was determined to be 3,243 pg/mL in HeLa treated and 139 pg/mL in untreated HeLa cell extract.

Interpolated concentrations of native c-Fos in RAW264.7 treated or untreated with starve 16 hours then TPA for 4 hours cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723957, Human/Mouse c-Fos Rabbit mAb [PSH17-92]
Detector: HA723958, Human/Mouse c-Fos Rabbit mAb [PSH17-93]

Interpolated concentration of native c-Fos was measured in duplicate at different sample concentrations and interpolated from the c-Fos standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Fos concentration was determined to be 1,907 pg/mL in RAW264.7 treated and 1,090 pg/mL in untreated RAW264.7 cell extract.

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

Capture: HA723957, Human/Mouse c-Fos Rabbit mAb [PSH17-92]
Detector: HA723958, Human/Mouse c-Fos Rabbit mAb [PSH17-93]

The concentrations of c-Fos were measured in duplicates, interpolated from the c-Fos 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).