SNAP-Cell^® TMR-Star

Product Information

This package contains 30 nmol of SNAP-Cell TMR-Star substrate, sufficient to make 10 ml of a 3 µM SNAP-tag fusion protein labeling solution.

The SNAP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on mammalian O⁶-alkylguanine-DNA alkyltransferase (AGT). SNAP-tag substrates are derivatives of benzylpurines and benzylchloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.

There are two steps to using this system: subcloning and expression of the protein of interest as a SNAP-tag fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of SNAP-tag fusion proteins is described in the documentation supplied with SNAP-tag plasmids. The labeling of the fusion proteins with the SNAP-tag substrate is described below.

Figure 1:

Figure 2: Excitation (dotted line) and emission spectra of SNAP-Cell TMR-Star after coupling to SNAP-tag in buffer at pH 7.5

Figure 3: Structure of SNAP-Cell TMR-Star (MW 677.1 g/mol)

This product is related to the following categories:

SNAP-tag Substrates Products ,

Protein Labeling Products

This product can be used in the following applications:

SNAP Cell,

Pulse Chase,

In vivo Imaging,

Receptor Internalization,

Protein Localization,

Protein Labeling Snap Clip

• Reagents Supplied

  The following reagents are supplied with this product:

  NEB https://www.neb.com/en/#	Component Name	Component https://www.neb.com/en/#	Stored at (°C)	Amount	Concentration
  S9105S     -20       SNAP-Cell® TMR-Star S9105SVIAL -20 1 x 30 nmol Not Applicable

• Properties & Usage

  Materials Required but not Supplied

  • Cells expressing SNAP-tag fusion proteins 
  • Tissue culture materials and media 
  • Transfection reagents 
  • Fluorescence microscope with suitable filter set 
  • DMSO

  Excitation

  554nm

  Emission

  580nm

• Product Notes

  1. Storage: SNAP-Cell TMR-Star should be stored at -20°C (long term) or at 4°C in the dark (short term, less than 4 weeks). Protect the substrate from light and moisture. With proper storage at -20°C the substrate should be stable for at least three years dry or 3 months dissolved in DMSO.
  2. While the majority of SNAP-tag substrates are benzylguanine derivatives, the SNAP-tag is also able to recognize chloropyrimidine derivatives. SNAP-Cell TMR-Star is one such example.

Protocols, Manuals & Usage

• Protocols

  1. Cellular Labeling (S9105)
  2. Labeling of Proteins in vitro (S9105)
  3. View the video "Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging" in the Journal of Visualized Experiments (JoVE)

• Application Notes

  • Simultaneous Fluorescent Labeling of Proteins in Living Cells

Tools & Resources

• Selection Charts

  • Comparison of SNAP-tag®/CLIP-tagâ„¢ Technologies to GFP
  • SNAP-tag® and CLIP-tagâ„¢ Substrate Selection Chart

FAQs & Troubleshooting

• FAQs

  1. Cellular Imaging and Analysis FAQs

• Troubleshooting

  • Labeling with SNAP-tag® Technology Troubleshooting Guide

• Tech Tips

  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).

  After the wash steps, incubate the intracellular labeling samples at 37°C, 5% CO₂ for 30 extra minutes to allow any unincorporated fluorophore to diffuse out of the cells.

Citations & Technical Literature

• Citations

  Product Citation Tool

  Additional Citations

  • Domoszlai T, Martincuks A, Fahrenkamp D, Schmitz-Van de Leur H, Küster A, Müller-Newen G (2014) Consequences of the disease-related L78R mutation for dimerization and activity of STAT3 J Cell Sci; 127(Pt 9), 1899-910. PubMedID: 24569879, DOI: https://www.neb.com/en/10.1242/jcs.137422

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.

• 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]

  • S9105S_v3

• 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]

  • S9105S_v3_10043113
  • S9105S_v3_10063236
  • S9105S_v3_10089443
  • S9105S_v3_10094849
  • S9105S_v3_10144354
  • S9105S_v3_10169901
  • S9105S_v3_10230853
  • S9105S_v3_10257649

• Safety Data Sheets

  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

  • SNAP-Cell^® TMR-Star

• 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

  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:
  Methods for Using O6-Alkylguanine-DNA-Alkyltransferases: US 7,939,284; US 8,367,361; EP 1 410 023; EP 2 037 271; EP 1 696 234; EP 2 211 177; JP 4195815; JP 4226053; CN 1295510; CN 1975422; and SG 100125.
  
  Mutants of O6-Alkylguanine-DNA-Alkyltransferases: US 7,888,090. EP 1720981 (pending).
  
  Pyrimidines Reacting With O6-Alkylguanine-DNA-Alkyltransferases: US 8,178,314. EP 1874738 (pending)