SNAP-Cell® Starter Kit

Companion Products

• dam^-/dcm^- Competent E. coli
• pSNAP-tag® (T7)-2 Vector
• Anti-SNAP-tag® Antibody (Polyclonal)
• SNAP-tag® Purified Protein
• SNAP-Cell® 505-Star
• SNAP-Cell® Oregon Green®
• SNAP-Cell® TMR-Star
• SNAP-Cell® Block
• SNAP-Cell® 430
• SNAP-Biotin®
• SNAP-Vista® Green

1. For long-term storage, all kit components should be stored at -20˚C. Plasmid solutions can be stored at 4˚C for up to one week. Undissolved dye and blocking substrates can be stored at 4˚C for up to 4 weeks protected from light and moisture. With proper storage at -20°C the substrates should be stable for at least three years dry or 3 months dissolved in DMSO.
2. NEB 10-beta Competent E. coli (High Efficiency) (NEB #C3019) is recommended for propagating and subcloning of the vector and control plasmid.

1. Cellular Labeling (E9100)
2. Use of SNAP-Cell Block with SNAP-Cell Substrates (E9100)
3. Labeling of Proteins in vitro (E9100)

• Simultaneous Fluorescent Labeling of Proteins in Living Cells

• Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
• Starter Kit Selection Chart

1. What is the SNAP-tag®?
2. How does it work?
3. How specific is the binding of substrate to the SNAP-tag®?
4. How does SNAP-tag labeling differ from using GFP fusion proteins?
5. What linker type and length would you recommend?
6. Can I clone my protein as a fusion to the N- or C-terminus of the tags?
7. What is the smallest peptide and biggest protein you have cloned as SNAP-tag fusions?
8. What is the solubility of SNAP-tag in insect and bacterial expression systems?
9. What competent cell strains does NEB suggest for expression in E. coli?
10. What competent cell E. coli strains are suitable for propagating SNAP-tag plasmids?
11. Can SNAP-tag fusions be purified and refolded from inclusion bodies?
12. Are substrates toxic to cells?
13. How does SNAP-tag affect localization of the fusion partner?
14. Can I use cell lines which express endogenous AGT?
15. How stable is the labeled protein in mammalian cells?
16. Are SNAP-tag substrates stable to fixation?
17. Can cells expressing SNAP-tag be fixed prior to labeling?
18. Can SNAP-tag be multiplexed with other protein labeling systems (GFP, Antibody)?
19. Can you use SNAP-tag for in vivo FRET?
20. Can cell-impermeable substrates be microinjected into cells, and how is the excess substrate exported?
21. Does the SNAP-tag labeling reaction work in yeast?
22. What happens to the fluorophore upon proteolysis?
23. What conditions are recommended for SNAP-tag labeling in vitro?
24. What conditions are incompatible with SNAP-tag labeling in vitro?
25. Can SNAP-tag fusion proteins be labeled in a cell lysate?
26. I have a compound that I would like to couple to a BG derivative. Where can I get advice?
27. What is the difference between SNAP-tag and ACP-tag?
28. What is the difference between SNAP- and CLIP-tag?
29. Cellular Imaging and Analysis FAQs

• Labeling with SNAP-tag® Technology Troubleshooting Guide

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.

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]

• E9100S_v1

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]

• E9100S_v1_10040415

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

• pSNAP_f Vector
• SNAP-Cell® 505-Star

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:
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).