Human IL-2 Receptor alpha Recombinant Rabbit Monoclonal Antibody [PS00-52] - BSA and Azide free (Capture)

The binding activity of IL-2 Receptor alpha (HA721280) with recombinant IL-2 Receptor alpha protein.

Immobilized recombinant IL-2 Receptor alpha protein at 1 μg/ml overnight at 4℃. Then blocked with 1xTBS/1%BSA for 1 hour at 37℃, and incubated with the primary antibody (HA721280) for 45min at 37℃. Then the plate was washed and incubated with 50 µl per well of Goat anti-Rabbit IgG-HRP for 0.5 hour at 37℃. 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.

Standard curve of IL-2 Receptor alpha matched pair antibodies:

Sandwich ELISA analysis of IL-2 Receptor alpha matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody HA721280 [PS00-52] diluted in carbonate/bicarbonate buffer, at a concentration of 4 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 1% BSA/PBST blocking buffer, and incubated with serial diluted recombinant IL-2 Receptor alpha protein starting from 2000 pg/ml to 31.25 pg/ml for 1 hour at 37℃. The plate was washed and incubated with 50 µl per well of detect antibody [PS01-03] (Biotin, 1:2,000) for 1 hour at 37℃. Then the plate was washed and incubated with 50 µl per well of Streptavidin-HRP for 0.5 hour at 37℃. 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 IL-2 Receptor alpha in human serum.

Capture: HA721280, Human IL-2 Receptor alpha Rabbit mAb [PS00-52]
Detector: HA721281, Human IL-2 Receptor alpha Rabbit mAb [PS01-03]

The concentrations of human IL-2 Receptor alpha measured in duplicate and interpolated from the human IL-2 Receptor alpha standard curve and corrected for sample dilution. Undiluted samples are human serum 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean human IL-2 Receptor alpha concentration was determined to be 1,304.63 pg/mL in human serum.