Catalog # S9220 was discontinued on December 15, 2022
CLIP-Cell™ Block (bromothenylcytosine, BTC) is a non-fluorescent compound that blocks the reactivity of the CLIP-tag™ in vitro and inside or on the surface of living cells.
It is cell permeable
It can be used to generate inactive controls in live cell labeling experiments performed with CLIP-tag fusion proteins
It reacts with CLIP-tag irreversibly, inactivating it for subsequent labeling steps
The CLIP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. CLIP-tag is a protein tag based on human O6-alkylguanine-DNAalkyltransferase (hAGT). CLIP-tag substrates are derivatives of benzylcytosine (BC). In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the reactive cysteine of CLIP-tag forming a stable thioether bond. Although CLIP-tag is based on the same protein as SNAP-tag®, the benzylcytosine substrates form a separate class of substrates, different from the benzylguanine substrates recognized by SNAP-tag. CLIP-tag and SNAP-tag can be used for orthogonal and complementary labeling of two proteins simultaneously in the same cells.
There are two steps to using this system: subcloning and expression of the protein of interest as a CLIP-tag fusion, and labeling of the fusion with the CLIP-tag substrate of choice. The use of BTC during the labeling of fusion proteins with CLIP-Cell substrates is described in this document.
Figure 1: Structure of CLIP-Cell Block (MW 286.1)
This product is related to the following categories:
Storage: CLIP-Cell Block should be stored at -20°C (long term) or at 4°C (short term). With proper storage at -20°C, CLIP-Cell Block is stable for at least three years dry or for three months when dissolved in DMSO.
For troubleshooting please refer to the instructions supplied with CLIP-Cell products as appropriate.
Please note that there is a constant turnover and resynthesis of proteins in the cell. After having blocked all existing CLIP-tag fusion proteins within the cell, new CLIP-tag fusion protein molecules may be synthesized in the meantime and may get labeled during incubation with a fluorescent CLIP-tag substrate. This will give the impression that the blocking was ineffective. In order to minimize these effects of protein synthesis and protein transport, cells may have to be treated with cycloheximide and incubation with the fluorescent CLIP-tag substrate may have to be performed at 4°C.
After addition of DMSO, pipette up and down at least 10-20 times and vortex vigorously for at least one full minute to ensure full dissolution of the substrate.
After diluting the substrate in complete medium, thoroughly pipette up and down at least 10 times to help reduce background.
Increasing substrate concentration and/or reaction time usually results in higher background and does not necessarily increase the signal-to-background ratio (SNR).
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.
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.
Licenses
Cellular Imaging and Analysis (i.e., SNAP and CLIP products)
Notice to Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938. For detailed information see our Terms of Use .
These patents and patent applications are owned by Covalys, or owned by the Ecole Polytechnique Fédérale de Lausanne (EPFL) and exclusively licensed to Covalys and NEB.
The products and/or their use may be covered by one or more of the following patents and patent applications: Labeling of Fusion Proteins with Synthetic Probes: US 8,227,602; US 8,623,627; and EP 2 049 499.
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