Target Synonym: DIF;TNF-alpha;TNFA;TNFSF2
Usage: This ELISPOT kit is designed for the detection of the frequency of human TNF-αsecreting cells.
Tested Sample Types: Suspension cell
Assay Time: 3 h 30 min
Test Principle: This assay kit uses the Sandwich-ELISA principle. The PVDF membrane at the bottom of the micro ELISPOT plate has been re-coated with anti-human TNF-α antibodies. During the experiment, cell suspension and stimulants are added first, and TNF-α secreted by the cells during the incubation process will bind to the coated antibodies. Then wash away the cells, add biotinylated anti-human TNF-α antibodies and alkaline phosphatase conjugated avidin sequentially. The biotinylated anti-human TNF-α antibodies will bind specifically to the TNF-α which has already bounded, then avidin will bind specifically to biotin, together forming the complex. After free components are washed away, add chromogenic substrate (BCIP/NBT), BCIP undergoes hydrolysis under the catalysis of alkaline phosphatase and reacts with NBT, forming insoluble spots with color dark blue to blue-purple. Professional ELISPOT plate reader software can be used to analyze the result, calculating the frequency of human TNF-α secreting cells.
Cross reactivity: This kit recognizes Human TNF-α in samples.No significant cross-reactivity or interference between Human TNF-α and analogues was observed.
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