Human Thymidine Kinase 1 / TK1 Recombinant Rabbit Monoclonal Antibody [PSH18-63] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human Thymidine Kinase 1 / TK1 matched pair antibodies

Capture: HA725239, Human Thymidine Kinase 1 / TK1 Rabbit mAb [PSH18-62]
Detector: HA725240, Human Thymidine Kinase 1 / TK1 Rabbit mAb [PSH18-63]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725239) 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 TK1 protein starting from 16,000 pg/ml to 0 pg/ml and detect antibody (HA725240, 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 TK1 in HT-29 cell extract and Jurkat cell extract:

Capture: HA725239, Human Thymidine Kinase 1 / TK1 Rabbit mAb [PSH18-62]
Detector: HA725240, Human Thymidine Kinase 1 / TK1 Rabbit mAb [PSH18-63]

The concentrations of TK1 were measured in duplicates, interpolated from the TK1 standard curve and corrected for sample dilution. Undiluted samples are HT-29 cell extract 250 µg/mL and Jurkat cell extract 250 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TK1 concentration was determined to be 17,292 pg/ml in HT-29 cell culture supernatant and 5,771 pg/ml in Jurkat cell extract.