cIAP1 Recombinant Rabbit Monoclonal Antibody [PSH06-71] - BSA and Azide free (Detector)

Sandwich ELISA analysis of human cIAP1 matched pair antibodies

Capture: HA723280, cIAP1 Rabbit mAb [PSH06-72]
Detector: HA723282, cIAP1 Rabbit mAb [PSH06-71]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723280) 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 Human cIAP1 protein (HA211301) starting from 8,000 pg/ml to 0 pg/ml and detect antibody (HA723282, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 cIAP1 matched pair antibodies

Capture: HA723281, cIAP1 Rabbit mAb [PSH02-10]
Detector: HA723282, cIAP1 Rabbit mAb [PSH20-70]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723281) 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 Human cIAP1 protein (HA211301) starting from 8,000 pg/ml to 0 pg/ml and detect antibody (HA723282, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 cIAP1 in THP-1 and TF-1 extract samples based on a 1,000 µg/ml extract load.

Capture: HA723280, cIAP1 Rabbit mAb [PSH06-72]
Detector: HA723282, cIAP1 Rabbit mAb [PSH06-71]

The concentrations of cIAP1 were measured in duplicates, interpolated from the cIAP1 standard curve and corrected for sample dilution. Undiluted samples are THP-1 extract 25% and TF-1 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean cIAP1 concentration was determined to be 1,712 pg/ml in THP-1 extract and 1,800 pg/ml in TF-1 extract.

Interpolated concentrations of native cIAP1 in THP-1 and TF-1 extract samples based on a 1,000 µg/mL extract load.

Capture: HA723281, cIAP1 Rabbit mAb [PSH02-10]
Detector: HA723282, cIAP1 Rabbit mAb [PSH06-71]

The concentrations of cIAP1 were interpolated from the cIAP1 standard curves and corrected for sample dilution. Undiluted samples are THP-1 extract 50% and TF-1 extract 50%. The mean cIAP1 concentration was determined to be 1,450 pg/ml in THP-1 extract and 2,235 pg/ml in TF-1 extract.

Interpolated concentrations of spiked cIAP1 in human cell culture media samples.

Capture: HA723280, cIAP1 Rabbit mAb [PSH06-72]
Detector: HA723282, cIAP1 Rabbit mAb [PSH06-71]

The concentrations of cIAP1 were measured in triplicates, interpolated from the cIAP1 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=3).

Interpolated concentrations of spiked cIAP1 in human cell culture media samples.

Capture: HA723281, cIAP1 Rabbit mAb [PSH02-10]
Detector: HA723282, cIAP1 Rabbit mAb [PSH06-71]

The concentrations of cIAP1 were interpolated from the cIAP1 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.