Mouse MDC/CCL22 Recombinant Rabbit Monoclonal Antibody [PSH15-76] - BSA and Azide free (Capture)

Sandwich ELISA analysis of mouse CCL22 matched pair antibodies

Capture: HA723780, Mouse MDC/CCL22 Rabbit mAb [PSH15-76]
Detector: HA723781, Mouse MDC/CCL22 Rabbit mAb [PSH15-77]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody MR240604-AA1341-1-1H9 or MR240604-AA1341-1-1C11 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 mouse CCL22 protein (HA211072) starting from 500 pg/ml to 0 pg/ml and detect antibody MR240604-AA1341-1-1E2 -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 CCL22 in mouse spleen cell supernatant treated or untreated with LPS for 48 hours.

Capture: HA723780, Mouse MDC/CCL22 Rabbit mAb [PSH15-76]
Detector: HA723781, Mouse MDC/CCL22 Rabbit mAb [PSH15-77]

Interpolated concentration of native CCL22 was measured in duplicate at different sample concentrations and interpolated from the CCL22 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CCL22 concentration was determined to be 238 pg/mL in mouse spleen treated cell supernatant, 54 pg/mL in neat untreated mouse spleen cell supernatant.

Interpolated concentrations of native CCL22 in J774A.1 cell supernatant treated or untreated with PHA and PMA for 72 hours.

Capture: HA723780, Mouse MDC/CCL22 Rabbit mAb [PSH15-76]
Detector: HA723781, Mouse MDC/CCL22 Rabbit mAb [PSH15-77]

Interpolated concentration of native CCL22 was measured in duplicate at different sample concentrations and interpolated from the CCL22 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CCL22 concentration was determined to be 554 pg/mL in neat J774A.1 treated supernatant, 17 pg/mL in neat untreated J774A.1 supernatant.

Interpolated concentrations of spiked CCL22 in cell culture media samples.

Capture: HA723780, Mouse MDC/CCL22 Rabbit mAb [PSH15-76]
Detector: HA723781, Mouse MDC/CCL22 Rabbit mAb [PSH15-77]

The concentrations of CCL22 were measured in duplicates, interpolated from the CCL22 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).