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Home Discontinued Products SNAP-Cell® Fluorescein

SNAP-Cell® Fluorescein

This product has been discontinued. SNAP-Cell 505 Star (NEB #S9103) and SNAP-Cell Oregon Green (NEB #S9104) can be used as alternatives.

Catalog # S9107 was discontinued on May 28, 2019 ;

SNAP-Cell^® Fluorescein is a green fluorescent substrate that can be used to label SNAP-tag^® fusion proteins inside living cells or in vitro. This substrate has limited photostability.

It is a cell-permeable substrate (BG-Fluorescein) based on diacetylfluorescein
Diacetylfuorescein is essentially non-fluorescent, but it becomes fluorescent inside the cell when it is hydrolyzed by non-specific esterases, yielding fluorescein
It is suitable for standard fluorescein filter sets
It has an excitation maximum at 500 nm and an emission maximum at 532 nm

Learn more about how this product is being used in the Product Citation Tool

Product Information
This package contains 50 nmol of SNAP-Cell Fluorescein substrate, sufficient to make 10 ml of a 5 µM SNAP-tag fusion protein labeling solution. This substrate has limited photostability. If this presents a problem, we recommend using SNAP-Cell 505, which has similar spectral characteristics but much greater photostability.

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 becomes covalently attached to the SNAP-tag.

There are two steps to using this system: sub-cloning 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 fusion proteins with the SNAP-tag substrate is described below.

Figure 1: Live CHO-K1 cells transiently transfected with pSNAP-H2B

Cells were labeled with SNAP-Cell Fluorescein (green) for 15 minutes.

Figure 2: Excitation (dotted line) and emission spectra of SNAP-Cell Fluorescein (fluorescent product of SNAP-Cell Fluorescein in cells) coupled to SNAP-tag in buffer at pH 7.5)

Figure 3: Structure of SNAP-Cell Fluorescein (MW 712.7g/mol)

Product Categories:

Discontinued Products
- 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
  500nm
  Emission
  532nm
  Storage Temperature
  -20°C
- Product Notes
  Storage:
  SNAP-CellFluorescein 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 andmoisture. With proper storage at -20°C the substrate should be stablefor at least 3 years dry or 3 months dissolved in DMSO.
Protocols, Manuals & Usage
- Protocols
  Cellular Labeling (S9107)
  
  View the video "Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging" in the Journal of Visualized Experiments (JoVE)
  
  Labeling of Proteins in vitro (S9107)
- Application Notes
  Simultaneous Fluorescent Labeling of Proteins in Living Cells S9107
- 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.)
  
  S9107Datasheet-Lot0081206
  
  S9107Datasheet-Lot0091401
  
  S9107Datasheet-Lot0091407
  
  S9107Datasheet-Lot0091602
Tools & Resources
- Selection Charts
  Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
  
  SNAP-tag®, CLIP-tag™ and ACP/MCP-tag Substrate Selection Chart
FAQs & Troubleshooting
- FAQs
  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
  
  Powered by Bioz See more details on Bioz
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
  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]
  
  S9107S_v1
  
  S9107S_v2
  
  S9107S_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]
  
  S9107S_v2_10018919
- 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® Fluorescein
- 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.)
  
  S9107Datasheet-Lot0081206
  
  S9107Datasheet-Lot0091401
  
  S9107Datasheet-Lot0091407
  
  S9107Datasheet-Lot0091602
- 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
  
  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.
  
  Substrates for O6-Alkylguanine-DNA Alkyltransferase: US 7,799,524; and JP 4976651.
  
  Mutants of O6-Alkylguanine-DNA-Alkyltransferases: US 7,888,090. EP 1720981 (pending).
  
  Specific Substrates for O6-Alkylguanine-DNA-Alkyltransferases: US 8,163,479. EP 1730298 (pending).

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