• My NEB
  • Print
  • PDF
  • SNAP-Surface® 649

    Description

    SNAP-Surface 649 is a photostable fluorescent substrate that can be used to label SNAP-tag® fusion proteins on cell surfaces and in solution. This substrate (BG-649) is based on the Dyomics dye DY-649P1 and is suitable for excitation with a 650 nm diode laser or use with Cy5 filter sets. It has an excitation maximum at 655 nm and emission maximum at 676 nm. This package includes 50 nmol of SNAP-Surface 649 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 SNAPtag substrate is described below.

    Excitation (dotted line) and emission spectra of SNAP-Surface 649 coupled to SNAP-tag in buffer at pH 7.5
    S9159a
    Structure of SNAP-Surface 649 (MW 1221.3 g/mol).

    Properties and 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 SNAPSurface 649.
    • Tissue culture materials and media
    • Transfection reagents
    • Fluorescence microscope with suitable filter set
    • DMSO

    Storage Temperature

    -20°C

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Cellular Protein Labeling (Cell Surface):
      The product is tested on cells expressing the target protein on the surface.  The surface target is labeled and visualized by fluorescence microscopy
    • In Vitro Protein Labeling:

      The product is tested in an in vitro protein labeling reaction. After incubation the labeled product is visualized on SDS-PAGE by fluorescent detection and verified by mass spectrometry.

    • Physical Purity (HPLC):
      The purity of the product is determined by HPLC analysis.

    Notes

    1. Storage: SNAP-Surface 649 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.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    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 info@neb.com or fill out the Technical Support Form for appropriate document.
    1. Cellular Imaging and Analysis FAQs
    1. Cellular Labeling (S9159)
    2. Labeling of Proteins in vitro (S9159)
    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)