Thermo Scientific Pierce Sulfo-NHS and NHS are chemical modification reagents for converting carboxyl groups to amine-reactive NHS esters for bioconjugation, crosslinking, labeling and immobilization methods.
Sulfo-NHS (N-hydroxysulfosuccinimide) and its uncharged analog NHS (N-hydroxysuccinimide) enable control and modification of carbodiimide crosslinking reactions involving activation of carboxylates (–COOH) for conjugation with primary amines (–NH2). Derivatives are easily synthesized by mixing the Sulfo-NHS with a carboxyl-containing molecule and a dehydrating agent such as the carbodiimide EDC (EDAC). The method is the basis for generating many types of protein labeling reagents, including amine-reactive fluorescent dyes, biotin affinity tags and pegylation compounds.
- Efficiency of EDC-mediated coupling is increased in the presence of NHS or Sulfo-NHS
- Amine-reactive NHS esters or Sulfo-NHS esters can be made with any carboxyl-containing molecule
- NHS derivatives of typical small molecules (e.g., biotin) are uncharged, organic-soluble (dissolve first in DMSO) and cell membrane-permeable
- Sulfo-NHS derivatives are usually directly water-soluble (can be added directly to physiologic buffers) and membrane-impermable (can be used for cell surface labeling)
- High-purity, crystalline NHS and Sulfo-NHS can be used to create high-quality activated derivatives
- Improve efficiency of EDC coupling reactions
- Convert carboxyls to amine-reactive Sulfo-NHS esters
- Crosslink proteins to carboxyl-coated beads or surfaces more efficiently
- Activate nanoparticles with amine-reactive Sulfo-NHS esters
|Chemical structures of NHS and Sulfo-NHS reagents.
Sulfo-NHS plus EDC (carbodiimide) crosslinking reaction scheme. Carboxyl-to-amine crosslinking using the carbodiimide EDC and sulfo-NHS. Addition of NHS or Sulfo-NHS to EDC reactions (bottom-most pathway) increases efficiency and enables molecule (1) to be activated for storage and later use. NHS-activation is the basis for synthesizing amine-reactive labeling reagents and crosslinking compounds.
Properties of NHS and Sulfo-NHS.
||Store at 4°C, protect from moisture, use only fresh solutions
Specifications for NHS and Sulfo-NHS:
We manufacture N-hydroxysulfosuccinimide to the highest possible specifications to produce the most specific bioconjugates, to ensure the integrity of your data and to provide you with the highest degree of consistency. Each lot of Sulfo-NHS is tested to meet the following minimum specifications:
- Purity – Greater than 95% by quantitative NMR (the highest standard for crosslinker purity);
average lot purity is greater than 99%
- Solubility – Sample dissolves at 2mg/mL in deionized water to yield a clear, colorless solution
- Identity – IR scan shows only peaks characteristic of N-hydroxysulfosuccinimide
Greater than 99% pure Sulfo-NHS. Prepared batches of Pierce Sulfo-NHS are tested by quantitative NMR against an internal standard to ensure purity and lot-to-lot consistency. Each peak in this typical NMR scan corresponds to a particular proton of Sulfo-NHS, the internal standard, or the solvents used in the NMR analysis.
- Grabarek, Z. and Gergely, J. (1990). Zero-length crosslinking procedure with the use of active esters. Anal Biochem 185:131-5.
- Staros, J.V., et al. (1986). Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. Anal Biochem 156:220-2.
- Lomant, A.J. and Fairbanks, G. (1976). Chemical probes of extended biological structures: synthesis and properties of the cleavable cross-linking reagent [35S] dithiobis(succinimidyl propionate). J Mol Biol 104:243-61.
- Cuatrecaseas, P. and Parikh, I. (1972). Adsorbents for affinity chromatography. Use of N-hydroxysulfosuccinimide esters of agarose. Biochemistry 11:291-9.
Crosslinker Application Guide -- search for recent literature references for this product
NHS and Sulfo-NHS FAQ
Overview of crosslinking
Chemistry of crosslinking
EDC (carbodiimide crosslinker)
Bioconjugate Techniques (book)
Crosslinkers at a glance (selection guide)
NHS-activated Biotinylation Reagents
NHS-activated Fluorescent Labeling Reagents
NHS-activated Agarose Resin
NHS-activated Magnetic Beads