Mouse MMP-9 Recombinant Rabbit Monoclonal Antibody [PSH15-80] - BSA and Azide free (Capture/Detector)

Sandwich ELISA analysis of Mouse MMP-9 matched pair antibodies

Capture: HA723784, Mouse MMP-9 Rabbit mAb [PSH15-79]
Detector: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723784) 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 MMP-9 protein (HA211045) starting from 6,000 pg/ml to 0 pg/ml and detect antibody (HA723785, 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 MMP-9 matched pair antibodies

Capture: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]
Detector: HA723787, Mouse MMP-9 Rabbit mAb [PSH15-81]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723785) 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 MMP-9 protein (HA211045) starting from 6,000 pg/ml to 0 pg/ml and detect antibody (HA723787, 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 MMP-9 in mouse serum samples.

Capture: HA723784, Mouse MMP-9 Rabbit mAb [PSH15-79]
Detector: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]

The concentrations of MMP-9 were measured in triplicates, interpolated from the MMP-9 standard curve and corrected for sample dilution. Undiluted samples are mouse serum 2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean MMP-9 concentration was determined to be 249.5 ng/ml in mouse serum.

Interpolated concentrations of native MMP-9 in mouse serum samples.

Capture: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]
Detector: HA723787, Mouse MMP-9 Rabbit mAb [PSH15-81]

The concentrations of MMP-9 were measured in duplicates, interpolated from the MMP-9 standard curve and corrected for sample dilution. Undiluted samples are mouse serum 2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MMP-9 concentration was determined to be 370.1 ng/ml in mouse serum.

Interpolated concentrations of native MMP-9 in mouse LPS unstimulated and stimulated J774A.1 cell culture supernatant.

Capture: HA723784, Mouse MMP-9 Rabbit mAb [PSH15-79]
Detector: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]

J774A.1 cells were stimulated with 2.5 µg/ml LPS or vehicle control and incubated for 1 day. The concentrations of MMP-9 measured in duplicate and interpolated from the MMP-9 standard curve and corrected for sample dilution. Undiluted samples are as follows: unstimulated 50% and stimulated 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MMP-9 concentration was determined to be 15,121 pg/ml in LPS stimulated J774A.1 cell culture supernatant and undetectable in the unstimulated J774A.1 control.

Interpolated concentrations of native MMP-9 in mouse LPS unstimulated and stimulated J774A.1 cell culture supernatant.

Capture: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]
Detector: HA723787, Mouse MMP-9 Rabbit mAb [PSH15-81]

J774A.1 cells were stimulated with 2.5 µg/ml LPS or vehicle control and incubated for 1 day. The concentrations of MMP-9 measured in duplicate and interpolated from the MMP-9 standard curve and corrected for sample dilution. Undiluted samples are as follows: unstimulated 50% and stimulated 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MMP-9 concentration was determined to be 21,179 pg/ml in LPS stimulated J774A.1 cell culture supernatant and undetectable in the unstimulated J774A.1 control.

Interpolated concentrations of spiked MMP-9 in mouse cell culture media samples.

Capture: HA723784, Mouse MMP-9 Rabbit mAb [PSH15-79]
Detector: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]

The concentrations of MMP-9 were measured in duplicates, interpolated from the MMP-9 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).

Interpolated concentrations of spiked MMP-9 in mouse cell culture media samples.

Capture: HA723785, Mouse MMP-9 Rabbit mAb [PSH15-80]
Detector: HA723787, Mouse MMP-9 Rabbit mAb [PSH15-81]

The concentrations of MMP-9 were measured in duplicates, interpolated from the MMP-9 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).