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EK0306: Nuclease ELISA Kit
Application: for the quantitative determination of nuclease proteins with the same amino acid sequence as MilliporeSigma Benzonase nuclease in cell culture supernatants.
This assay is specific for Benzonase nuclease and its alternative enzymes.
Detection range: 46.88 pg/mL - 3000.00 pg/mL
Sensitivity: 23.44 pg/mL
Precision: CV < 10%
Kit components for 96-well plate ELISA for Benzonase nuclease and its alternative enzymes (1x 96-well microplate):
1 Pre-coated Microplate: 96 well polystyrene microplates coated with monoclonal antibody specific for nuclease;
2 Detection Antibody: a horseradish peroxidase (HRP)- conjugated polyclonal antibody against nuclease; 1 vial (20 ul)
3 Nuclease Standard: 10,000 ng/ml nuclease protein; 1 vial (50 ul)
4 Assay Diluent: 1x PBS with 1% BSA; 1 bottle (60 ml)
5 Wash Buffer: 10x Concentrate- PBS with detergent; 1 bottle (60 ml)
6 Color Reagent: tetramethylbenzidine (TMB); 1 bottle (20 ml)
7 Stop Solution: 0.5 N hydrochloric acid solution; 1 bottle (20 ml)
8 Product Insert: product description, assay protocol, material data sheet; 1 book.
Background
MilliporeSigma Benzonase nuclease and its alternative enzymes including Dr. Nuclease are a genetically engineered endonuclease derived from Serratia marcescens that can degrade all forms of DNA and RNA including double-stranded, single-stranded, circular, or linear RNA and DNA completely to 5'-monophosphate oligonucleotides 2-5 bases in length over a wide range of reaction conditions and is therefore widely used to completely digest and remove nucleic acid residues from biological samples.
This kit uses sandwich ELISA to determine the concentration of Benzonase nuclease in the test sample. The capture anti-nuclease monoclonal antibody is pre-coated on the 96-well plate. Nuclease standard or test sample is added to the pre-coated 96-well plate and will specifically bind to the capture antibody. Then the HRP-conjugated nuclease detection antibody is added to bind the immune complex forming the antibody-antigen-detection antibody-HRP complex. The plate is rinsed to remove extra HRP conjugated nuclease detection antibody and then TMB is added for chromatic changes. The amplitude of the color change is proportional to the amount of nuclease that specifically binds to the plate. The reaction is stopped with the addition of stop solution and the absorbance is measured at 450 nm. The sample nuclease concentration is calculated from the standards titration curve.
Nuclease:
Recombinant Dr. Nuclease
RNase A Enzyme
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