Efficient replacement for Tobacco Acid Pyrophosphatase
Cap0 and Cap1 are removed with equal efficiency
Suitable for 5′ RLM-RACE and RNA-seq
1 µl will decap 20 µg of a 1.7 kb mRNA in 15 minutes
Product Information
mRNA Decapping Enzyme catalyzes the removal of 7-methylguanosine cap (m7G) from 5´ end of mRNA, producing 5′ monophosphate and releasing m7GDP(1). mRNA Decapping Enzyme is capable of decapping mRNAs of various lengths and removes both Cap0 and Cap1 structures with similar efficiency. mRNA Decapping Enzyme also converts 5′ triphosphate ends to 5′ monophosphate, albeit with reduced efficiency. 5′ monophosphorylated RNA can be exploited in a variety of downstream applications, including 5′ RNA Ligase-mediated RACE, RNA-seq, and 5′-> 3′ exonuclease digestion.
Comparison of mRNA Decapping Enzyme and Tobacco Acid Pyrophosphatase Decapping Activity.
(Panel A) A 500 nM solution of 25 nucleotide 5′ capped RNA was treated with either mRNA Decapping Enzyme (MDE) or competitor Tobacco Acid Pyrophosphatase (TAP). Both reactions were carried out in the same reaction volume and using each enzyme’s appropriate reaction buffer. While the concentration of a competitor’s TAP used is unknown, this comparison reveals 0.62 units MDE to have a similar level of activity relative to 1 µl of TAP. Decapping reaction products were analyzed by capillary electrophoresis. Note that in the negative control reaction there is a contaminating amount of triphosphorylated RNA of ~10% in the synthetic substrate used.(Panel B) To test the impact of introducing length and termini heterogeneity into the reaction, 500 ng of total RNA for Jurkat cells was added while maintaining all other conditions unchanged. Decapping by MDE was unaffected, whereas TAP activity was markedly diminished, indicating that MDE is the more robust decapping enzyme under these test conditions.
Product Source
mRNA Decapping Enzyme from S.pombe is expressed as a His-tagged fusion in E. coli.
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
M0608S
-20
mRNA Decapping Enzyme
M0608SVIAL
-20
1 x 0.02 ml
100,000 units/ml
mRNA Decapping Enzyme Reaction Buffer
B0608SVIAL
-20
1 x 1 ml
10 X
Properties & Usage
Unit Definition
One unit is defined as the amount of mRNA Decapping Enzyme required to convert 50% of a 500 nM m7G-capped substrate to a 5´-monophosphorylated form in a total reaction volume of 20 µl in 1 hour at 37˚C.
Reaction Conditions
1X mRNA Decapping Enzyme Reaction Buffer
Incubate at 37°C
1X mRNA Decapping Enzyme Reaction Buffer
50 mM Tris-HCl
50 mM Ammonium Chloride
5 mM MgCl2
1 mM DTT
0.1% Poloxamer 188
(pH 7.5 @ 25°C)
Storage Buffer
300 mM NaCl
10 mM Tris-HCl
0.1 mM DTT
50% Glycerol
pH 7.4 @ 25°C
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 & Change Notifications
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]
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]
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.
Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Please review and update your order accordingly. If you have any questions, please contact Customer Support at freezers@neb.com or 1-800-632-5227 x 8.
Choose your country
North America
Europe
Asia-Pacific
Session Expired
Your session has expired
You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.
Please sign in to continue shopping.
Institution Changed
Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.
Sign in to your NEB account
To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.
Powered by Bioz
