Biotin Conjugated Human CKMB Recombinant Rabbit Monoclonal Antibody [PSH11-56] - Detector

Sandwich ELISA analysis of human CKMB matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723348) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 CKMB protein (HA210777) starting from 80000 pg/ml to 0 pg/ml and detect antibody (HA723350B, 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 CKMB in rat heart and rat skeletal muscle extract samples based on a 1000 µg/ml extract load.

Interpolated concentration of native CKMB was measured in duplicate at different sample concentrations and interpolated from the CKMB standard curves. The mean CKMB concentration was determined to be 62,193 pg/mL in rat heart tissue extract. There was no detectable signal in rat skeletal muscle extract.

Interpolated concentrations of spiked CKMB in cell culture media samples.

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