T7 RNA Polymerase ELISA kit

T7 RNA polymerase uses double-stranded DNA containing the T7 promoter sequence (5'-TAATACGACT CACTATAG*-3') as a template and NTP as a substrate to synthesize RNA complementary to the reverse single-stranded DNA downstream of the promoter. Double-stranded linear blunt-end or 5' protruding end DNA can be used as a substrate template for T7 RNA polymerase, so linear plasmids and PCR products can be used as templates for in vitro RNA synthesis.
This kit uses the experimental principle of double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to detect the residual amount of T7 RNA polymerase. T7 RNA polymerase standards and samples to be tested are added to the ELISA plate pre-coated with anti-T7 RNA polymerase antibodies. Then, diluted biotin-labeled T7 RNA polymerase detection antibodies are added, and finally Streptavidin-HRP (SA-HRP) is added to form an antibody + antigen + antibody-Biotin + SA-HRP complex. After washing the plate, TMB color development solution is added for color development. TMB is converted from colorless to blue under the catalysis of HRP enzyme and finally converted to yellow under the action of stop solution. The depth of yellow is positively correlated with the amount of T7 RNA polymerase detected in the sample.

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