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        How does SSTK Assay work?
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        How does SSTK Assay work?

        1.The ECL dot-blot assay was performed according to the manufacturer’s protocol. Blood sample of 2 ml was collected in the morning between 7 and 9 a.m. from persons who did not take any breakfast.

        2. The drawn venous blood in non-heparin tube was stored for 2–3 h at room temperature (RT) and centrifuged at 4,000 rpm for 10 min and then stored at -20°C before analysis. Three microliters of serum was directly applied onto an Amersham Hybond TM ECL TM membrane.

        3. The membrane was blocked in Tris-buVered saline (TBS) solution with 6% non-fat milk for 1 h and then primary anti-TK1 antibody was added and incubated at room temperature (RT) for 2h. After incubation with a biotinylated secondary antibody for 40 min at RT, the membrane was incubated in TBS solution with avidin/streptavidin-HRP, followed by addition of the substrate ECL.

        4. The light intensity of the spots on the membrane was detected by a CIS-2 imaging system. From the light intensities of the TK1 standard of known concentrations, the light intensities of the serum TK1 spots were re-calculated and expressed as pM.

        5.The ECL Dot Blot assay technology amplifies the signal, enabling the assay to measure thymidine kinase 1 (TK1) with high sensitivity.

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