Human cIAP2 Recombinant Rabbit Monoclonal Antibody [PSH09-39] - BSA and Azide free (Capture/Detector)

Sandwich ELISA analysis of human cIAP2 matched pair antibodies.

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723098) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Human cIAP2 Protein (HA210619) starting from 20000 pg/ml to 0 pg/ml and detect antibody (HA723099, 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 human cIAP2 matched pair antibodies.

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723099) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Human cIAP2 Protein (HA210619) starting from 20000 pg/ml to 0 pg/ml and detect antibody (HA723100, 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 cIAP2 in Raji, Ramos and SH-SY5Y extract samples based on a 1000 µg/ml extract load.

Interpolated concentration of native cIAP2 was measured in duplicate at different sample concentrations and interpolated from the cIAP2 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean cIAP2 concentration was determined to be 4,595 pg/mL in Raji and 927 pg/mL in Ramos cell extract, There was no detectable signal in SH-SY5Y cell extract.

Interpolated concentrations of native cIAP2 in Raji, Ramos and SH-SY5Y extract samples based on a 1000 µg/ml extract load.

Interpolated concentration of native cIAP2 was measured in duplicate at different sample concentrations and interpolated from the cIAP2 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean cIAP2 concentration was determined to be 1,390 pg/mL in Raji and 561 pg/mL in Ramos cell extract, There was no detectable signal in SH-SY5Y cell extract.

Interpolated concentrations of spiked cIAP2 in cell extract samples.

The concentrations of cIAP2 were measured in duplicates, interpolated from the cIAP2 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell extract 0.2mg/ml. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Interpolated concentrations of spiked cIAP2 in cell culture media samples.

The concentrations of cIAP2 were measured in duplicates, interpolated from the cIAP2 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).