Biotin Conjugated Human/Rat/Monkey BDNF Recombinant Rabbit Monoclonal Antibody [PSH27-09] - Detector

Sandwich ELISA analysis of Human BDNF matched pair antibodies

Capture: HA724669, Human/Rat/Monkey BDNF Rabbit mAb [PSH27-08]
Detector: HA724670, Human/Rat/Monkey BDNF Rabbit mAb [PSH27-09]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724669) diluted in carbonate/bicarbonate buffer, at a concentration of 2 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 BDNF protein (HA211599) starting from 1,000 pg/mL to 0 pg/mL and detect antibody (HA724670, 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 BDNF in human, cynomolgus monkey and rat samples.

Capture: HA724669, Human/Rat/Monkey BDNF Rabbit mAb [PSH27-08]
Detector: HA724670, Human/Rat/Monkey BDNF Rabbit mAb [PSH27-09]

Interpolated concentration of native BDNF was measured in duplicate at different sample concentrations.The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean BDNF concentration was determined to be 9,552 pg/mL in human serum, 2,193 pg/mL in cynomolgus monkey serum and 1,462 pg/mL in rat serum.

Interpolated concentrations of spiked BDNF in cell culture media samples.

Capture: HA724669, Human/Rat/Monkey BDNF Rabbit mAb [PSH27-08]
Detector: HA724670, Human/Rat/Monkey BDNF Rabbit mAb [PSH27-09]

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