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Home Discontinued Products pCLIP_f-H2B Control Plasmid

pCLIP_f-H2B Control Plasmid

Catalog # N9218 was discontinued on January 05, 2015 ;

Product Information
This control plasmid contains the gene encoding the Histone H2B protein cloned upstream of the CLIP_f coding sequence in pCLIP_f as a fusion to the N- terminus of the CLIP-tag. Histone H2B is a member of the core histones that package DNA in the nucleus. The H2B-CLIP_f fusion protein gives nuclear fluorescence when labeled with CLIP-Cell™ substrates. The full sequence and map for pCLIP_f-H2B can be downloaded.

The CLIP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The CLIP-tag is a small protein 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 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_ffusion, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of the CLIP_f-H2B fusion protein is described in this document. The labeling of fusion proteins with CLIP-tag substrates is described in the instructions supplied with CLIP-tag substrates.

Figure 1: Live CHO-K1 cells transiently transfected with pCLIP_f-H2B. Cells were labeled with CLIP-Cell™ 505 (green) for 30 minutes and counterstained with Hoechst 33342 (blue).

DNASU is a central repository for plasmid clones and collections that may also be helpful.

Product Categories:

Discontinued Products
- Properties & Usage
  Materials Required but not Supplied
  
  Tissue 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-H2B 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-H2B (N9218)
- Application Notes
  Simultaneous Fluorescent Labeling of Proteins in Living Cells N9218
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  N9218Datasheet-Lot0061206
  
  N9218Datasheet-Lot0011012
  
  N9218Datasheet-Lot0021411
Tools & Resources
- Selection Charts
  Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
  
  SNAP-tag®/CLIP-tag® Cloning Vector and Plasmid 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.
  
  pCLIP_f-H2B Control Plasmid
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  N9218Datasheet-Lot0061206
  
  N9218Datasheet-Lot0011012
  
  N9218Datasheet-Lot0021411
- 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.
  For Research Use Only: 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.
  
  FuGENE® is a registered trademark of Roche.

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