Human CXCL13 Recombinant Rabbit Monoclonal Antibody [PSH07-05] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human CXCL13 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722791) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 human CXCL13 protein (HA211065) starting from 2000 pg/ml to 0 pg/ml and detect antibody (HA722792, Biotin, 0.3 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 Human CXCL13 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722791) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 human CXCL13 protein (HA211065) starting from 2000 pg/ml to 0 pg/ml and detect antibody (HA722794, Biotin, 0.3 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.

☑ Cell treatment (CT)

Interpolated concentrations of native CXCL13 in human THP-1 supernatant treated or untreated with Hu-IFN-r and LPS for 24 hours.

Interpolated concentration of native CXCL13 was measured in duplicate at different sample concentrations and interpolated from the CXCL13 standard curves. Undiluted samples were 20% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CXCL13 concentration was determined to be 8,902 pg/mL in neat THP-1 treated supernatant, undetectable in untreated THP-1 supernatant.

☑ Cell treatment (CT)

Interpolated concentrations of native CXCL13 in human THP-1 supernatant treated or untreated with Hu-IFN-r and LPS for 24 hours.

Interpolated concentration of native CXCL13 was measured in duplicate at different sample concentrations and interpolated from the CXCL13 standard curves. Undiluted samples were 20% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CXCL13 concentration was determined to be 10,653 pg/mL in neat THP-1 treated supernatant, undetectable in untreated THP-1 supernatant.

Interpolated concentrations of spiked CXCL13 in human cell culture media samples.

The concentrations of CXCL13 were measured in duplicates, interpolated from the CXCL13 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 CXCL13 in human cell culture media samples.

The concentrations of CXCL13 were measured in duplicates, interpolated from the CXCL13 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).