Biotin Conjugated Neuropeptide Y Recombinant Rabbit Monoclonal Antibody [PSH16-89] - Detector

Sandwich ELISA analysis of Human NPY matched pair antibodies

Capture: HA723884, Neuropeptide Y Rabbit mAb [PSH16-88]
Detector: HA723885, Neuropeptide Y Rabbit mAb [PSH16-89]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723884) 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 Neuropeptide Y protein (HA211303) starting from 750 pg/ml to 0 pg/ml and detect antibody (HA723885, 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 NPY in mouse hippocampus and rat hippocampus tissue extract samples based on a 1000 µg/ml extract load.

Capture: HA723884, Neuropeptide Y Rabbit mAb [PSH16-88]
Detector: HA723885, Neuropeptide Y Rabbit mAb [PSH16-89]

Interpolated concentration of native NPY was measured in duplicate at different sample concentrations and interpolated from the NPY standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean NPY concentration was determined to be 1,192 pg/mL in mouse hippocampus and 565 pg/mL in rat hippocampus tissue extract.

Interpolated concentrations of native NPY in human SH-SY5Y cell supernatant treated or untreated with TPA for 24 hours.

Capture: HA723884, Neuropeptide Y Rabbit mAb [PSH16-88]
Detector: HA723885, Neuropeptide Y Rabbit mAb [PSH16-89]

Interpolated concentration of native NPY was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean NPY concentration was determined to be 735 pg/mL in neat SH-SY5Y treated supernatant and undetectable in untreated SH-SY5Y supernatant.

Interpolated concentrations of spiked NPY in cell culture media samples.

Capture: HA723884, Neuropeptide Y Rabbit mAb [PSH16-88]
Detector: HA723885, Neuropeptide Y Rabbit mAb [PSH16-89]

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