Human TRAF1 Recombinant Rabbit Monoclonal Antibody [PSH14-28] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human TRAF1 matched pair antibodies

Capture: HA723618, Human TRAF1 Rabbit mAb [PSH14-27]
Detector: HA723619, Human TRAF1 Rabbit mAb [PSH14-28]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723618) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 TRAF1 protein (HA211289) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723619, 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 TRAF1 in Raji and HEK-293 extract samples based on a 1,000 µg/ml extract load.

Capture: HA723618, Human TRAF1 Rabbit mAb [PSH14-27]
Detector: HA723619, Human TRAF1 Rabbit mAb [PSH14-28]

The concentrations of TRAF1 were measured in duplicates, interpolated from the TRAF1 standard curve and corrected for sample dilution. Undiluted samples are Raji extract 20% and HEK-293 extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TRAF1 concentration was determined to be 10,599 pg/ml in Raji extract and undetectable in HEK-293 extract.

Interpolated concentrations of spiked TRAF1 in human cell culture media samples.

Capture: HA723618, Human TRAF1 Rabbit mAb [PSH14-27]
Detector: HA723619, Human TRAF1 Rabbit mAb [PSH14-28]

The concentrations of TRAF1 were measured in duplicates, interpolated from the TRAF1standard 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).