Biotin Conjugated Human/Mouse/Rat FGF1 Recombinant Rabbit Monoclonal Antibody [PSH18-35] - Detector

Sandwich ELISA analysis of Human FGF1 matched pair antibodies

Capture: HA723987, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-34]
Detector: HA723988, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-35]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723987) 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 FGF1 protein (HA211079) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723988, 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 FGF1 in U-87 MG cell extract samples based on a 1,000 µg/ml extract load.

Capture: HA723987, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-34]
Detector: HA723988, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-35]

The concentrations of FGF1 were measured in duplicates, interpolated from the FGF1 standard curve and corrected for sample dilution. Undiluted samples are U-87 MG cell extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FGF1 concentration was determined to be 337 pg/ml in U-87 MG cell extract.

Interpolated concentrations of native FGF1 in mouse brain tissue extract and rat heart tissue extract samples based on a 1,000 µg/ml extract load.

Capture: HA723987, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-34]
Detector: HA723988, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-35]

The concentrations of FGF1 were measured in duplicates, interpolated from the FGF1 standard curve and corrected for sample dilution. Undiluted samples are mouse brain tissue extract 5% and rat heart tissue extract 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FGF1 concentration was determined to be 3,420 pg/ml in mouse brain tissue extract and 7,048 pg/ml in rat heart tissue extract.

Interpolated concentrations of spiked FGF1 in human cell culture media samples.

Capture: HA723987, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-34]
Detector: HA723988, Human/Mouse/Rat FGF1 Rabbit mAb [PSH18-35]

The concentrations of FGF1 were measured in duplicates, interpolated from the FGF1 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).