Mouse SAA Recombinant Rabbit Monoclonal Antibody [PSH25-06] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Mouse SAA matched pair antibodies

Capture: HA724500, Mouse SAA Rabbit mAb [PSH25-06]
Detector: HA724501, Mouse SAA Rabbit mAb [PSH25-07]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724500) 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 Mouse SAA1 protein (HA211238) starting from 5,000 pg/mL to 0 pg/mL and detect antibody (HA724501, 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 SAA matched pair antibodies

Capture: HA724500, Mouse SAA Rabbit mAb [PSH25-06]
Detector: HA724501, Mouse SAA Rabbit mAb [PSH25-07]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724500) 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 Mouse SAA1 protein (HA211238) and Recombinant Mouse SAA2 protein starting from 5,000 pg/mL to 0 pg/mL and detect antibody (HA724501, 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 SAA in mouse serum.

Capture: HA724500, Mouse SAA Rabbit mAb [PSH25-06]
Detector: HA724501, Mouse SAA Rabbit mAb [PSH25-07]

Interpolated concentration of native SAA was measured in duplicate at different sample concentrations and interpolated from the SAA standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean SAA concentration was determined to be 5.06 μg/mL in mouse serum.

Interpolated concentrations of spiked SAA1 in cell culture media samples.

Capture: HA724500, Mouse SAA Rabbit mAb [PSH25-06]
Detector: HA724501, Mouse SAA Rabbit mAb [PSH25-07]

The concentrations of SAA1 were measured in duplicates, interpolated from the SAA1 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).