BG-PEG-NH2

Description

BG-PEG-NH2 is an amine-terminated building block for the one-step synthesis of SNAP-tag substrates from NHS esters or other activated carboxyl esters on labels or surfaces. This compound allows you to make custom SNAP-tag substrates for labeling SNAP-tag fusion proteins for a wide range of applications. BG-PEG-NH2 is particularly useful for the synthesis of SNAP-tag substrates with affinity ligands or solid surfaces.

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 fusion proteins with the SNAP-tag substrate is described in this document.
S9150a
Figure 1. Structure of BG-PEG-NH2 (MW 488.50 g/mol)

Notes

  1. Dry BG-PEG-NH2 should be stored at -20°C.
  2. BG-PEG-NH2 is soluble in anhydrous N,N-dimethylformamide (DMF) and dimethyl sulfoxide (DMSO). Choose a solvent volume that will be compatible with the final use. Heating at 37°C, vortexing and/or sonication will aid dissolution.
  3. Unused BG-PEG-NH2 substrate dissolved in DMF or DMSO should be stored at -20°C.
  4. Surfaces such as Biacore® CM5 chips have been modified with BG-PEG-NH2 so that they covalently immobilize SNAP-tag fusion proteins.  Surface activation is performed using standard amino-coupling procedures (exposure to NHS and EDC).  Refer to Biacore® CM5 chips manufacturer's protocol for detailed instruction.

References

  1. Kindermann, M. et al. (2003). JACS . 125, 7810-7811.
  2. Sielaff, I. et al. (2003). ChemBiochem. 7, 194-202.
  3. Kufer, S.K. et al. (2005). Eur. Biophys. J. 35, 72-78.
  4. Huber, W. et al. (2004). Anal. Biochem. 333, 280-288.

FAQs

  1. Cellular Imaging and Analysis FAQs

Protocols

  1. Labeling SNAP-tag Fusion Proteins with BG-substrates (S9150)
  2. Reaction Conditions for Chemical coupling (S9150)

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.

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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. Further information regarding NEB product quality can be found here.
  • Physical Purity (HPLC):
    The purity of the product is determined by HPLC analysis.

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]

Safety Data Sheet

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

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.