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Micrococcal Nuclease (MNase)


Shear or cleave nucleic acids in cell lysates for chromatin IP (ChIP) assays.

Micrococcal Nuclease (MNase)

Thermo Scientific Micrococcal Nuclease (MNase) is suitable for removing nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments.

This micrococcal nuclease is a stable liquid form of the enzyme derived from Staphylococcus aureus. Micrococcal nuclease (MNase) exhibits exo- and endo-5'-phosphodiesterase activities against DNA and RNA. This enzyme digests double-stranded, single-stranded, circular and linear nucleic acids. The highest activity is toward single-stranded nucleic acid substrates with preference for AT- or AU-rich regions. Enzymatic activity occurs at pH 7 to 10 and is strictly dependent on calcium for digestion of RNA and DNA substrates. Micrococcal Nuclease is suitable for removing nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments.

Highlights:

  • Enzyme digests nucleic acids (DNA and RNA)
  • Effective for double-stranded, single-stranded, circular and linear nucleic acids
  • Active in neutral to alkaline buffer conditions containing divalent calcium ions
  • Supplied at ≥100 units/µL in 10mM Tris-HCl pH 7.5, 50mM NaCl, 1mM EDTA, 50% glycerol

Properties of Micrococcal Nuclease (MNase):

  • Product is a liquid at -20°C. Keep enzyme on ice during use.
  • Micrococcal nuclease is dependent on Ca++ for activity. Avoid calcium chelators, such as EGTA, in reaction buffers.
  • Enzyme is active at pH 7 to 10 with a salt concentration less than 100mM. Optimal enzyme activity occurs at 37°C; however, the enzyme is active at room temperature. Monovalent metal ions, such as Na+ and K+, will decrease activity. EGTA or heating to 65°C for 10 minutes will inactivate the enzyme.

Generalized Protocol for use of Micrococcal Nuclease (MNase):

  1. Dilute sample in 50mM Tris-HCl pH 8.0, 5mM CaCl2
  2. If sample only contains nucleotides (i.e., no protein), add BSA at a final concentration of 0.1mg/mL.
  3. Add micrococcal nuclease and incubate reaction at 37°C until the nucleotides are degraded.
  4. Optional: Stop reaction by adding EGTA, pH 8.0 to a final concentration of 20mM.

Specifications of Micrococcal Nuclease (Part No. 88216):

  • Visual: Clear, colorless liquid free of insoluble material.
  • Units: ≥100 units/µL
  • Additional Information: 1 unit is defined as the amount of enzyme required to produce an increase in absorbance at 260nm of 0.001/min/mL at 25°C of highly polymerized DNA.

Related Resources:

Review of chromatin IP (ChIP) assays

Related Products:

Pierce ChIP Kit
Subcellular Fractionation Kit
DNase I


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
88216 Micrococcal Nuclease
Formulation: Enzyme in 10mM Tris-HCl pH 7.5, 50mM NaCl, 1mM EDTA, 50% glycerol
150┬ÁL Product Instructions for product #88216 Micrococcal Nuclease MSDS for product #88216 Micrococcal Nuclease Certificate of Analysis for product #88216 Micrococcal Nuclease $113.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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