Product Class: Other

T4 PDG (T4 Endonuclease V)
NEBU recombinant 37 No ralbumin

The large size of this product has been discontinued.

Product Introduction

  • T4 pyrimidine DNA glycosylase
  • Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities
  • The N-glycosylase activity releases cis-syn cyclobutane pyrimidine dimers, including T^T, T^C and C^C, generating an AP site. The AP lyase activity cleaves an AP site via β-elimination, creating a 1 nucleotide DNA gap with 3'-α, β-unsaturated aldehyde and 5'-phosphate termini.
  • Also known as T4 Endonuclease V
Catalog # Size Concentration
M0308S 2000.0 units 10000 units/ml

Product Information

Description

T4 PDG (pyrimidine dimer glycosylase) has both DNA glycosylase and APlyase activity. The 16 kd protein recognizes cis-syn-cyclobutane pyrimidine dimers caused by UV irradiation. The enzyme cleaves the glycosyl bond of the 5´ end of the pyrimidine dimer and the endonucleolytic activity cleaves the phosphodiester bond at the AP site.

Product Source

Purified from an E. coli strain carrying a plasmid encoding T4 denV gene.
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 that catalyzes the conversion of 0.5 µg of UV irradiated supercoiled pUC19 DNA to > 95% nicked plasmid in a total reaction volume of 20 µl in 30 minutes at 37°C. Nicking is assessed by agarose gel electrophoresis. Irradiated plasmid contains an average of 3-5 pyrimidine dimers.

Reaction Conditions

1X T4 PDG Reaction Buffer
Supplement with 100 µg/ml Recombinant Albumin, Molecular Biology Grade
Incubate at 37°C

1X T4 PDG Reaction Buffer
100 mM NaCl
1 mM DTT
1 mM EDTA
25 mM Na2HPO4
(pH 7.2 @ 25°C)

Storage Buffer

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

Heat Inactivation

No

Unit Assay Conditions

1X T4 PDG Reaction Buffer containing 0.5 µg of UV irradiated supercoiled pUC19 DNA, supplemented with 100 µg/ml Recombinant Albumin in a 20 µl reaction.

Application Features

  • DNA damage studies
  • Single cell gel electrophoresis (comet assay)

Product Notes

  1. Warm buffer to room temperature, as it precipitates at 4°C.
  2. The enzyme has 100% activity in NEBuffer 4 (NEB #B7004) and 75% activity in NEBuffers 1-3 (NEB #B7001, NEB #B7002, NEB #B7003).
  3. For best results, incubation time should be 30 minutes or less.
  4. After the reaction, addition of SDS to a final concentration of 0.5%, or 1µl phenol, in 6X Loading dye to the sample before running the gel will improve electrophoresis by dissociating the protein from the DNA.

References

  1. Higgins, K.L. and Lloyd, R.S. (1987). Mutation Research. 183, 117-121.

Protocols, Manuals & Usage

Protocols

  1. Protocol for T4 PDG (M0308)

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the activity of T4 PDG in other NEBuffers, including rCutSmart?
  2. What is the molecular weight of T4 PDG?
  3. Is T4 PDG a tagged protein?
  4. Can the T4 PDG be used in the comet assay?
  5. Does T4 PDG remove the damaged base?
  6. What type of damaged DNA does T4 PDG recognize?

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.

Specification Change Notifications

Recombinant Albumin (NEB #B9200 at 20 mg/ml) replaced Purified BSA (NEB #B9001 at 10 mg/ml).

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 busdev@neb.com.

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.

Licenses

Nucleic acid-based aptamers for use with reverse transcriptases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the WarmStart® RTx Reverse Transcriptase for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com. 

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