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Home Discontinued Products pCLIP_f-Cox8A Control Plasmid

pCLIP_f-Cox8A Control Plasmid

Catalog # N9217 was discontinued on January 01, 2013 ;

Product Information
This control plasmid contains the gene encoding the Cytochrome C oxidase, subunit 8-2 (COX8-2) protein cloned upstream of the CLIP_f coding sequence in pCLIP_f, as a fusion to the N- terminus of the CLIP-tag. Cytochrome C oxidase is located in the inner mitochondrial membrane and is the terminal enzyme of the respiratory chain. The COX8-2-CLIP_f fusion protein gives mitochondrial fluorescence when labeled with CLIP-Cell™ substrates. The full sequence and map for pCLIP_f-Cox8A can be downloaded.

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-DNA-alkyltransferase (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.

pCLIP_f-Cox8A contains an improved version of CLIP-tag, termed CLIP_f. CLIP_f displays faster kinetics in in vitro labeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics.

There are two steps to using this system: sub cloning and expression of the protein of interest as a CLIP_f fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expressing the CLIP_f-Cox8A fusion protein is described in this document. The labeling of the fusion proteins with CLIP-tag substrates is described in the instructions supplied with CLIP-tag substrates.

Figure 1: Live HEK293 cells transiently transfected with pCLIP_f-Cox8A. Cells were labeled with CLIP-Cell TMR-Star (red) for 30 minutes and counterstained with Hoechst 33342 (blue).

Product Categories:

Discontinued Products
- Properties & Usage
  
  Materials Required but not Supplied
  Cell culture media and reagents
  Mammalian cell lines
  Transfection reagents
  CLIP-tag substrates
  Storage Temperature
  -20°C
  Sequence Files
  Fasta GenBank
- Product Notes
  Storage: pCLIP_f-Cox8A is supplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of 0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. For long-term storage -20°C is recommended.
Protocols, Manuals & Usage
- Protocols
  Expression of CLIPf-Cox8A (N9217)
- Application Notes
  Simultaneous Fluorescent Labeling of Proteins in Living Cells N9217
Tools & Resources
- Selection Charts
  Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
  
  SNAP-tag®/CLIP-tag® Cloning Vector Selection Chart
- Web Tools
  DNA Sequences and Maps Tool
FAQs & Troubleshooting
- FAQs
  Cellular Imaging and Analysis FAQs
- Troubleshooting
  Labeling with SNAP-tag® Technology Troubleshooting Guide
- Tech Tips
  
  If you generate more plasmid DNA by bacterial transformation, we recommend isolating the plasmid DNA using an endotoxin-free plasmid prep kit prior to transfection into mammalian cells.
Quality, Safety & Legal
- Quality Control Assays
  
  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.
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at [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.
  Licenses
  
  Notice To Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT PURPOSES ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938.
  
  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.
  
  The products and/or their use may be covered by one or more of the following patents and patent applications:
  
  7,939,284 (Methods for Using O6-Alkylguanine-DNA-Alkyltransferases)
  
  7,888,090 (Mutants of O6-Alkylguanine-DNA-Alkyltransferases)
  
  8,163,479 Specific Substrates for O6-Alkylguanine-DNA-Alkyltransferases)
  
  8,178,314 (Pyrimidines Reacting With O6-Alkylguanine-DNA-Alkyltransferases)
  
  PCT/EP2007/057597 (Labeling of Fusion Proteins with Synthetic Probes)
  
  EP07117800 (Drug Delivery)
  
  EP07117802 (Drug Delivery)
  
  EP07120288 (GTPase-Transient Protein Protein Interactions)
  
  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 CMV promoter is covered under U.S. Patent No. 5,385,839 and its use is permitted for research purposes only. Any other use of the CMV promoter requires a license from the University of Iowa Research Foundation, 214 Technology Innovation Center, Iowa City, IA 52242.
  No other use is authorized, including without limitation Commercial Use. Commercial Use means any and all uses of the product and derivatives by a party for monetary or other consideration including, but not limited to:
  
  (1) product manufacture;
  (2) providing of a service, information or data;
  (3) resale of the product, derivatives or components;
  (4) use of the product, derivatives or components by therapeutic, diagnostic or prophylactic clinical or commercial purposes.
  
  Information regarding any other license may be obtained by contacting Business Development and Licensing, New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938. Tel: 978-927-5054, Fax: 978-921-1350.

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