Product Class: Other

SNAP-Surface® Alexa Fluor® 488

Product Introduction

SNAP-Surface® Alexa Fluor® 488 is a photostable green fluorescent substrate that can be used to label SNAP-tag® fusion proteins in solution or on the surface of living cells.

  • This is a cell impermeable substrate based on BG-Alexa Fluor 488
  • It is suitable for standard fluorescein filter sets
  • It has an excitation maximum at 496 nm and emission maximum at 520 nm
Catalog # Size Concentration
S9129S 50.0 nmol

Product Information

Description

This package includes 50 nmol of SNAP-Surface Alexa Fluor 488 substrate, sufficient to make 10 ml of a 5 µM SNAP-tag fusion protein labeling solution.

The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is based on mammalian O6-alkylguanine-DNA-alkyltransferase (AGT). SNAP-tag
substrates are derivatives of benzylguanines 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: Live HEK293 cells transiently transfected with pSNAP-ADRβ2 and pSNAP-H2B

Figure 1: Live HEK293 cells transiently transfected with pSNAP-ADRβ2 and pSNAP-H2B

Cells were sequentially labeled with SNAP-Surface Alexa Fluor 488 (green) and SNAP-Cell TMR-Star (red) for 15-30 minutes.
Figure 2: Excitation (dotted line) and emission (full line) spectra of SNAP-Surface Alexa Fluor® 488 coupled to SNAP-tag in buffer at pH 7.5
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 Surface Products,
In vivo Imaging,
Pulse Chase,
Receptor Internalization,
Protein Localization,
Protein Labeling Snap Clip

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Materials Required but not Supplied

  • Cells expressing SNAP-fusion proteins. Proteins of interest can be expressed with the SNAP-tag as either an N-or a C-terminal fusion, but note that the tag needs to be exposed to the extracellular surface of the plasma membrane for labeling with SNAP-Surface Alexa Fluor-488.
  • Tissue culture materials and media
  • Transfection reagents
  • Fluorescence microscope with suitable filter set
  • DMSO

Excitation

496nm

Emission

520nm

Product Notes

  1. Storage: SNAP-Surface AlexaFluor 488 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 three years dry or 3 months dissolved in DMSO.

Protocols, Manuals & Usage

Protocols

  1. Cellular Labeling (S9129)
  2. Labeling Proteins in vitro (S9129)
  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

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Cellular Imaging and Analysis FAQs

Troubleshooting

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).

Quality, Safety & Legal

Quality Assurance Statement

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]

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]

Safety DataSheets

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

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

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.

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.

This product is provided under an intellectual property license from Life Technologies Corporation. The purchase and/or transfer of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research for directed covalent labeling of proteins conducted by the buyer (whether the buyer is an academic or for-profit entity), according to the product instructions. The buyer must not use the product or its components (1) in manufacturing; (2) to provide a service, information, or data; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008, USA or outlicensing@lifetech.com.

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). 

Trademarks

SNAP-Surface and SNAP-Cell are trademarks owned by New England Biolabs, Inc.

Alexa-Fluor and Oregon Green are trademarks owned by Life Technologies Corporation

Alexa-Fluor is a trademark owned by Life Technologies Corporation

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