Product Class: Other

BG-GLA-NHS

Product Introduction

BG-GLA-NHS is an amine-reactive building block for the one-step synthesis of SNAP-tag® substrates from amine-containing precursors including proteins, peptides or oligonucleotides.

  • This building block allows you to make custom SNAP-tag substrates for labeling SNAP-tag fusion proteins
Catalog # Size Concentration
S9151S 2.0 mg

Product Information

Description

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 protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). SNAP-tag substrates are fluorophores, biotin or beads conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker. 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: 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 the fusion proteins with the SNAP-tag substrate is described in this document.
9147a_thumb
Figure 1. Structure of BG-GLA-NHS (MW 481.5 g/mol)
 
This product is related to the following categories:
Building Blocks Products,
SNAP-tag Substrates Products ,
Protein Labeling Products,
This product can be used in the following applications:
Protein Labeling Snap Clip,
Cell Imaging

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

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

Properties & Usage

Reaction Conditions

Coupling BG-GLA-NHS to Proteins
NHS coupling to proteins is often performed using an excess of NHS ester. Depending on the number of lysine residues in the protein, it may be necessary to stop the labeling reaction before it reaches completion to avoid damaging the protein or causing it to unfold. A typical coupling involves reacting the NHS ester with exposed primary amines on the surface of the proteins to form an amide bond. The reaction can be carried out at pH 7-9 (typically pH 8.3), temperatures from 4–37°C and incubation times ranging from 30 min to overnight. Care must be taken to avoid primary amine containing buffers such as Tris or ammonium ions.

Coupling BG-GLA-NHS to Amine-Containing Compounds
A typical procedure involves equimolar quantities of NHS ester, amine to be coupled and triethylamine in DMF at 30°C overnight. The purification strategy will depend on the label used. Good results have been obtained with both HPLC and silica gel chromatography

Product Notes

  1. BG-GLA-NHS is supplied in lyophilized form and packaged under argon. The drySNAP-tag building block substrate should be stored at -20°C.
  2. BG-GLA-NHS is soluble in anhydrous DMF or DMSO. Choose a solvent volume thatwill be compatible with the final use. Vortex and centrifuge briefly to removeany non-dissolved solids.
  3. NHS esters readily hydrolyze in moist environments. Experiments should bedesigned to maximize the usage of BG-GLA-NHS. Storage of unused BG-GLA-NHSsolubilized in DMF or DMSO should be avoided. If unavoidable, we recommendstoring at -20°C for the minimum time period.

References

  1. Howland, S.W. et al. (2008). J. Immunother. 31, 607-619.

Protocols, Manuals & Usage

Protocols

  1. Labeling SNAP-tag Fusion Proteins with BG-substrates (S9151)
  2. Reaction Conditions for Chemical Coupling (S9151)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Cellular Imaging and Analysis FAQs

Troubleshooting

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 [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.

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.

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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