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Endonuclease IV

Catalog # Concentration Size List Price Quantity Your Price
M0304L 10000 units/ml 5000 units $503.00
$452.70
M0304S 10000 units/ml 1000 units $123.00
$110.70
Catalog # Size List Price Your Price
M0304L 5000 units $503.00
$452.70
M0304S 1000 units $123.00
$110.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • DNA AP endonuclease
  • Catalyzes the cleavage of DNA phosphodiester backbone at AP sites via hydrolysis leaving a 1 nucleotide gap with 3'-hydroxyl and 5' deoxyribose phosphate (dRP) termini
  • Also has 3'-diesterase activity which can remove 3' phosphate, 3'-α, β-unsaturated aldehyde, phosphoglycoaldehyde, and other 3' blocking groups
Featured Videos
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  • Monarch_MiniprepProtocol_VideoThumb
    What are endonucleases and their applications?
Endonuclease IV can act on a variety of oxidative damage in DNA (1). The enzyme is apurinic/apyrimidinic (AP) endonuclease that will hydrolyse intact AP sites in DNA. AP sites are cleaved at the first phosphodiester bond that is 5´ to the lesion leaving a hydroxyl group at the 3´ terminus and a deoxyribose 5´-phosphate at the 5´ terminus. The enzyme also has a 3´ -diesterease activity and can release phosphoglycolaldehyde, intact deoxyribose 5-phosphate and phosphate from the 3´ end of DNA.
Product Source
An E.coli strain which carries the cloned Endo IV gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Endonuclease IV M0304SVIAL -20 1 x 0.1 ml 10,000 units/ml
  NEBuffer™ 3 B7003SVIAL -20 1 x 1.25 ml 10 X
  Endonuclease IV M0304LVIAL -20 1 x 0.5 ml 10,000 units/ml
  NEBuffer™ 3 B7003SVIAL -20 1 x 1.25 ml 10 X
Application Features
  • Single cell gel electrophoresis (Comet assay)(2,3,4)
  • Alkaline elution (5)
  • Alkaline unwinding (6)

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 AP site* in a total reaction volume of 10 µl in 1 hour at 37°C.

*An AP site is created by treating 10 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C.

Reaction Conditions

1X NEBuffer™ 3
Incubate at 37°C

1X NEBuffer™ 3
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
0.15% Triton® X-100
pH 7.4 @ 25°C

Heat Inactivation

85°C for 20 minutes

Unit Assay Conditions

1X NEBuffer 3 containing 10 pmol of fluorescently labeled oligonucleotide duplex in a total reaction volume of 10 μl.

Companion Products
Materials Sold Separately
Notes
  • Recommended Dilution for the Comet Assay: 1.104 to 1:105(2,3,4,7). For a protocol please visit: http://cometassay.com
  • The Endo IV optimum buffer is 1X NEBuffer 3 (NEB #B7003). The enzyme has 100% activity in NEBuffer 2 (NEB #B7002) and 25% activity in CutSmart Buffer (NEB #B7204). The use of Endo IV in NEBuffer 1 (NEB #B7001) is not recommended. 
  • The enzyme is also active in ThermoPol® Reaction Buffer (NEB #B9004).
References
  • Levin, J. et al. (1988). J. Gen. Physiol. . 33, 349-362.
  • Singh, N. et al. (1961). Experimental Cell  Research . 175, 184-191.
  • Collins, A. etal. (1993). Carcinogenesis . 14, 1733-1735.
  • Collins, A. et al. (1996). Environmental Health Perspectives . 104, 465-469.
  • Pflaum, M. et al. (1998). Free Rad. Res. 29, 585-594.
  • Hartwig, A. et al. (1996). Toxicology Letters. 88, 85-90.
  • Marks, K., New England Biolabs, Inc. unpublished observations.
Quality Control Assay
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 Data Sheets
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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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