Thrombospondin 1 Recombinant Rabbit Monoclonal Antibody [PSH27-64] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human Thrombospondin 1 protein matched pair antibodies

Capture: HA724708, Thrombospondin 1 Rabbit mAb [PSH27-63]
Detector: HA724709, Thrombospondin 1 Rabbit mAb [PSH27-64]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724708) 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 Thrombospondin 1 protein (HA211369) starting from 10,000 pg/mL to 0 pg/mL and detect antibody (HA724709, Biotin, 0.05 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 Thrombospondin 1 in HUVEC cell culture supernatant.

Capture: HA724708, Thrombospondin 1 Rabbit mAb [PSH27-63]
Detector: HA724709, Thrombospondin 1 Rabbit mAb [PSH27-64]

HUVEC cells were cultured in media with or without FBS for 1 day. The concentrations of Thrombospondin 1 were measured in duplicates, interpolated from the Thrombospondin 1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombospondin 1 concentration was determined to be 59,247 pg/mL in HUVEC with FBS cell culture supernatant and 6,494 pg/mL in HUVEC without FBS cell culture supernatant.

Interpolated concentrations of native Thrombospondin 1 in human serum and monkey serum samples.

Capture: HA724708, Thrombospondin 1 Rabbit mAb [PSH27-63]
Detector: HA724709, Thrombospondin 1 Rabbit mAb [PSH27-64]

The concentrations of Thrombospondin 1 were measured in duplicates, interpolated from the Thrombospondin 1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombospondin 1 concentration was determined to be 3,228 ng/mL in human serum and 1,338 ng/mL in cynomolgus monkey serum.

Interpolated concentrations of native Thrombospondin 1 in bovine serum samples.

Capture: HA724708, Thrombospondin 1 Rabbit mAb [PSH27-63]
Detector: HA724709, Thrombospondin 1 Rabbit mAb [PSH27-64]

The concentrations of Thrombospondin 1 were measured in duplicates, interpolated from the Thrombospondin 1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombospondin 1 concentration was determined to be 3,813 ng/mL in bovine serum.

Interpolated concentrations of spiked Thrombospondin 1 in human cell culture media samples.

Capture: HA724708, Thrombospondin 1 Rabbit mAb [PSH27-63]
Detector: HA724709, Thrombospondin 1 Rabbit mAb [PSH27-64]

The concentrations of Thrombospondin 1 were measured in duplicates, interpolated from the Thrombospondin 1 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media without FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).