Biotin Conjugated Mouse Adiponectin Recombinant Rabbit Monoclonal Antibody [PSH14-83] - Detector

Sandwich ELISA analysis of mouse Adiponectin matched pair antibodies

Capture: HA723679, Mouse Adiponectin Rabbit mAb [PSH14-82]
Detector: HA723680, Mouse Adiponectin Rabbit mAb [PSH14-83]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723679) 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 Mouse Adiponectin protein (HA210973) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723680, 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.

Sandwich ELISA analysis of mouse Adiponectin matched pair antibodies

Capture: HA723682, Mouse Adiponectin Rabbit mAb [PSH14-84]
Detector: HA723680, Mouse Adiponectin Rabbit mAb [PSH14-83]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723682) 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 Mouse Adiponectin protein (HA210973) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723680, 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 Adiponectin in mouse samples.

Capture: HA723679, Mouse Adiponectin Rabbit mAb [PSH14-82]
Detector: HA723680, Mouse Adiponectin Rabbit mAb [PSH14-83]

Interpolated concentration of native Adiponectin was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Adiponectin concentration was determined to be 32 ug/mL in mouse serum and 49 ng/ml in mouse lung cell supernatant. There was no detectable signal in NIH/3T3 cell culture supernatant.

Interpolated concentrations of native Adiponectin in mouse samples.

Capture: HA723682, Mouse Adiponectin Rabbit mAb [PSH14-84]
Detector: HA723680, Mouse Adiponectin Rabbit mAb [PSH14-83]

Interpolated concentration of native Adiponectin was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Adiponectin concentration was determined to be 31 ug/mL in mouse serum and 47 ng/ml in mouse lung cell supernatant. There was no detectable signal in NIH/3T3 cell culture supernatant.

Interpolated concentrations of spiked Adiponectin in cell culture media samples.

Capture: HA723679, Mouse Adiponectin Rabbit mAb [PSH14-82]
Detector: HA723680, Mouse Adiponectin Rabbit mAb [PSH14-83]

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

Interpolated concentrations of spiked Adiponectin in cell culture media samples.

Capture: HA723682, Mouse Adiponectin Rabbit mAb [PSH14-84]
Detector: HA723680, Mouse Adiponectin Rabbit mAb [PSH14-83]

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