Biotin Conjugated S100 beta Recombinant Rabbit Monoclonal Antibody [PSH15-25] - Detector

Sandwich ELISA analysis of Human S100B matched pair antibodies

Capture: HA723728, Human S100B Rabbit mAb [SC57-02]
Detector: HA723729, Human S100B Rabbit mAb [PSH15-25]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723728) 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 S100B protein (HA211252) starting from 6,000 pg/ml to 0 pg/ml and detect antibody (HA723729, 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 S100B in SK-MEL-28 and T-47D extract samples based on a 1,000 µg/ml extract load.

Capture: HA723728, Human S100B Rabbit mAb [SC57-02]
Detector: HA723729, Human S100B Rabbit mAb [PSH15-25]

The concentrations of S100B were measured in triplicates, interpolated from the S100B standard curve and corrected for sample dilution. Undiluted samples are SK-MEL-28 extract 0.25% and T-47D extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean S100B concentration was determined to be 1,536 ng/ml in SK-MEL-28 extract and undetectable in T-47D extract.

Interpolated concentrations of spiked S100B in serum-free media samples.

Capture: HA723728, Human S100B Rabbit mAb [SC57-02]
Detector: HA723729, Human S100B Rabbit mAb [PSH15-25]

The concentrations of S100B were measured in duplicates, interpolated from the S100B standard curves and corrected for sample dilution. Undiluted samples are as follows: serum-free media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).