Human Ki67 Recombinant Rabbit Monoclonal Antibody [SR00-02] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human Ki67 matched pair antibodies

Capture: HA724536, Human Ki67 Rabbit mAb [PSH25-46]
Detector: HA724537, Human Ki67 Rabbit mAb [SR00-02]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724536) 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 Recombinant Human Ki67 protein (HA211578) starting from 8,000 pg/mL to 0 pg/mL and detect antibody (HA724537, 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 Ki67 in A431 and HeLa cell extract samples based on a 1,000 µg/mL extract load.

Capture: HA724536, Human Ki67 Rabbit mAb [PSH25-46]
Detector: HA724537, Human Ki67 Rabbit mAb [SR00-02]

The concentrations of Ki67 were measured in duplicates, interpolated from the Ki67 standard curve and corrected for sample dilution. Undiluted samples are A431 cell extract 50% and HeLa cell extract 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Ki67 concentration was determined to be 13,077 pg/mL in A431 cell extract and 39,701 pg/mL in HeLa cell extract.

Interpolated concentrations of spiked Ki67 in human cell culture media samples.

Capture: HA724536, Human Ki67 Rabbit mAb [PSH25-46]
Detector: HA724537, Human Ki67 Rabbit mAb [SR00-02]

The concentrations of Ki67 were measured in duplicates, interpolated from the Ki67 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).