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UltraNuclease, also known as a non-restrictive endonuclease, broad-spectrum nuclease, is a non-specific endonuclease derived from Serratia Marcescen, which hydrolyzes internal phosphodiester bonds between any nucleotides in nucleic acids to produce 5'-monophosphate oligonucleotides of 2-5 bases in length. It can degrade DNA and RNA in various forms (double-stranded, single-stranded, linear, circular, native or denatured) under a very broad range of conditions (6 M Urea, 0.1 M Guanidine HCl, 0.4% Triton X-100, 0.1% SDS, 1 mM EDTA, 1 mM PMSF) and is widely used to remove nucleic acids from biological products. UltraNuclease can also be removed by corresponding methods subsequently.
This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residues of denatured and non-denatured UltraNuclease. First, coat the microplate with an anti-UltraNuclease rabbit polyclonal antibody to form a solid-phase antibody. Second, add UltraNuclease standard and test sample to the solid-phase antibody microplate, then add biotin labeled Anti-UltraNuclease polyclonal antibody, and finally, add horseradish peroxidase-labeled streptavidin (SA-HRP) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, TMB substrate was added to the complex to observe color reaction after washing the complex. TMB is converted into blue under the catalysis of HRP enzyme and finally converted into yellow in the presence of acid, and the shade of color is positively correlated with the amount of UltraNuclease in the sample.
The detection quantification range of this kit is 0.047-3 ng/mL; the lower detection limit is 23.5 pg/mL.
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