Product Class: Other

Endonuclease V
NEB4 cloned at NEB recombinant 37 65 Heat

The large size of this product has been discontinued.

Product Introduction

  • DNA endonuclease
  • Catalyzes the cleavage of the second DNA phosphodiester backbone 3' to deoxyinosine, leaving a 3'-OH and 5' - phosphate
  • Endonuclease V activity, to a lesser extent, is also reported on AP site, urea, mismatches, hairpins, loops, and pseudo-Y structures
Catalog # Size Concentration
M0305S 250.0 units 10000 units/ml

Product Information

Description

Endonuclease V is a repair enzyme found in E. coli that recognizes deoxyinosine, a deamination product of deoxyadenosine in DNA. Endonuclease V, often called deoxyinosine 3´ endonuclease, recognizes DNA containing deoxyinosines (paired or not) on double-stranded DNA, single-stranded DNA with deoxyinosines and to a lesser degree, DNA containing abasic sites (ap) or urea, base mismatches, insertion/deletion mismatches, hairpin or unpaired loops, flaps and pseudo-Y structures. It is believed that Endonuclease V needs another protein to repair the DNA, as it does not remove the deoxyinosine or the damaged bases (1,2,3).

Endonuclease V cleaves the second phosphodiester bonds 3´ to the mismatch of deoxyinosine (2), leaving a nick with 3´-hydroxyl and 5´-phosphate (4).

Product Source

An E. coli strain containing a gene fusion of the Endo V gene and the gene coding for the maltose binding protein (MBP). The fusion protein is purified to near homogeneity and is active as a fusion. The protein contains 223 amino acids and has a molecular weight of 24.9 kDa (5).
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34 mer oligonucleotide duplex containing a single deoxyinosine site* in a total reaction volume of 10 μl in 1 hour at 37°C.

* A deoxyinosine site is synthetically prepared with a dl in the middle of one strand of a 34 mer oligonucleotide duplex.

Reaction Conditions

1X NEBuffer™ 4
Incubate at 37°C

1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
250 mM NaCl
0.1 mM EDTA
50% Glycerol
0.15% Triton® X-100
1 mM DTT
200 µg/ml BSA
pH 8 @ 25°C

Heat Inactivation

65°C for 20 minutes

Unit Assay Conditions

1X NEBuffer 4 containing 10 pmol of fluorescently labeled oligonucleotide duplex in a total volume of 10 μl.

Application Features

  • Mismatch Cleavage
  • Cleavage of oligonucleotides containing deoxyinosines and a weaker affinity for oligonucleotides containing base mismatches (5)

Product Notes

  1. Endonuclease V has full activity in NEBuffer 1 (NEB #B7001), NEBuffer 2 (NEB #B7002), and CutSmart (NEB #B7204). It has 75% activity in NEBuffer 3 (NEB #B7003).

References

  1. Yao, M. and Kow, Y.W. (1996). J. Biol. Chem. 271, 30672-30673.
  2. Yao, M. and Kow, Y.W. (1995). J. Biol. Chem.. 270, 28609-28616.
  3. Yao, M. et. al. (1994). J. Biol. Chem.. 269, 31390-31396.
  4. He, B., Qing, H. and Kow, Y.W. (2000). Mutat. Res.. 459, 109-114.
  5. Yao, M. and Kow, Y.W. (1994). J. Biol. Chem.. 269, 31390-31396.
  6. Marks, K. and Landry D., New England Biolabs, Inc. unpublished observation.

Protocols, Manuals & Usage

Protocols

  1. Protocol for Endonuclease V (M0305)

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the molecular weight of Endonuclease V?
  2. What substrate is used to test Endonuclease V activity?
  3. What is the activity of Endonuclease V in the different NEBuffers?
  4. How well does Endonuclease V cleave at U:A sites?
  5. Can Endonuclease V cleave the loop in a stem-loop DNA structure?
  6. Can a nick generated by Endonuclease V activity be repaired with DNA ligase?
  7. Can Endonuclease V recognize and nick mismatches?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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