Ultra Nuclease Assay Kit (ELISA)

UltraNuclease is a genetically engineered broad-spectrum nuclease originating from Serratia Marcescens. It can degrade all forms of DNA and RNA, including single-stranded, double-stranded, linear, natural and denatured nucleic acids, and digest them into 5′-oligonucleotide monophosphate of 3-5 base in length with no base-specific recognition. Therefore, it is the most ideal enzyme preparation for removing nucleic acid.

Principle of the Procedure

This UltraNuclease ELISA kit is a sandwich ELISA to be performed in a microplate format. Sample potentially containing endonuclease are incubated in microtiter plate wells which have been pre-coated with an affinity purified anti-endonuclease capture antibody. After incubation and a washing step in which unbound components are removed, an enzyme-conjugated, anti-endonuclease detector antibody is added. The immunological reactions result in the formation of a sandwich complex of solid phase antibody-endonuclease-enzyme labeled antibody. After a final washing step, a substrate solution is added to the wells and reacted, resulting in color development. The optical density is measured photometrically and is proportional to the analyte concentration present in the wells. The endonuclease concentration in unknown samples can be calculated based on the corresponding standard curve.

Kit Component

1. ELISA microplate….……….………………….8×12 wells
2. Enzyme conjugate (100×)…………………0.15mL×1
3. Dilution buffer…..………..……………………30mL×1
4. Substrate solution……………..….…………6mL×2
5. Stop solution…….……….……..……………..6mL×1
6. Wash concentrate (10×)…………………..50mL×1
7. Standard concentrate……………………..0.5mL×1
8. Sealing film………….…….….………………..4 films
9. Manual……………………..……………………..1 copy

Storage and Stability

1. Unopened kit should be stored at 2℃ to 8℃ for stability until the expiration date printed on the kit.

2. Opened reagents should be stored at 2℃ to 8℃ and are stable for 6 weeks.

Equipment required

1. Microplate reader spectrophotometer with dual wavelength capability at 450 and 650nm. (If your plate reader does not provide dual wavelength analysis you may read at just the 450nm wavelength.)

2. Microtiter plate rotator (400-600 rpm).

Reagent Preparation

1. Bring all reagents to room temperature (23℃±2℃) before use.

2. Prepare washing buffer (1×) by diluting 50mL of the Wash concentrate (10×) with 450mL of deionized or distilled water.

3. Enzyme conjugate (1×) : Dilute to work concentrate at 1:99 by dilution buffer before use.

4. Dilute the Standard concentrate (1000ng/mL) with dilution buffer to 20ng/mL, 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.63ng/mL, 0.31ng/mL, 0ng/mL. Standards (20ng/mL to 0ng/mL) are prepared according to the following dilution scheme:

1. Remove nucleic acids wrapped around the surface of virus particles, to promote virus release and purification.
2. Remove the exogenous nucleic acids in vaccine products, to lower the risk of residual toxicity of nucleic acid and improve product safety.
3. Reduce feed viscosity caused by nucleic acid during drug purification, accelerate the process of purification treatment, and increase protein recovery yield.
4. Remove nucleic acids in gene therapy, to obtain purified adeno-associated virus.
5. Used in sample preparation during column chromatography, electrophoresis and western blotting, to improve resolution and recovery rate.
6. Prevent cell clustering
7.