Biotin Conjugated Mouse CXCL16 Recombinant Rabbit Monoclonal Antibody [PSH17-72] - Detector

Sandwich ELISA analysis of Mouse CXCL16 matched pair antibodies

Capture: HA723940, Mouse CXCL16 Rabbit mAb [PSH17-71]
Detector: HA723941, Mouse CXCL16 Rabbit mAb [PSH17-72]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723940) 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 CXCL16 protein (HA211059) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723941, 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 CXCL16 in mouse samples.

Capture: HA723940, Mouse CXCL16 Rabbit mAb [PSH17-71]
Detector: HA723941, Mouse CXCL16 Rabbit mAb [PSH17-72]

Interpolated concentration of native CXCL16 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CXCL16 concentration was determined to be 4,600 pg/mL in RAW264.7 cell supernatant and 1,220 pg/ml in 4T1 cell supernatant. There was no detectable signal in B16-F1 cell culture supernatant.

Interpolated concentrations of spiked CXCL16 in cell culture media samples.

Capture: HA723940, Mouse CXCL16 Rabbit mAb [PSH17-71]
Detector: HA723941, Mouse CXCL16 Rabbit mAb [PSH17-72]

The concentrations of CXCL16 were measured in duplicates, interpolated from the CXCL16 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).