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NHS and Sulfo-NHS

Improve the efficiency of EDC coupling or create amine-reactive reagents!

Pierce Sulfo-NHS (N-hydroxysulfosuccinimide)

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

Product Details:

N-Hydroxysuccinimide; NHS
N-Hydroxysulfosuccinimide; Sulfo-NHS
Chemical structures of NHS and Sulfo-NHS reagents.


Reaction diagram for crosslinking with EDC and Sulfo-NHS

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.

Learn more:


Properties of NHS and Sulfo-NHS.
  NHS Sulfo-NHS
Molecular formula C4H5NO3 C4H4NNaO6S
Molecular weight 115.09 217.13
CAS Number 6066-82-6 106627-54-7
Storage conditions 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
Purity of Sulfo-NHS tested by quantitative NMR

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.


  1. Grabarek, Z. and Gergely, J. (1990). Zero-length crosslinking procedure with the use of active esters. Anal Biochem 185:131-5.
  2. Staros, J.V., et al. (1986). Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. Anal Biochem 156:220-2.
  3. 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.
  4. Cuatrecaseas, P. and Parikh, I. (1972). Adsorbents for affinity chromatography. Use of N-hydroxysulfosuccinimide esters of agarose. Biochemistry 11:291-9.

Product References:

Crosslinker Application Guide -- search for recent literature references for this product

Related Resources:

NHS and Sulfo-NHS FAQ
Overview of crosslinking
Chemistry of crosslinking

Related Products:

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

Ordering Information
Product # Description Pkg. Size Instructions MSDS CofA Price
24500 NHS
Formulation: N-Hydroxysuccinimide; powder solid
Sufficient For: 43440mL of typical 1X (5mM) reaction solution, or approx. 12500 uses at 2mg reagent per use
25g Product Instructions for product #24500 NHS MSDS for product #24500 NHS Certificate of Analysis for product #24500 NHS $44.00

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24510 Sulfo-NHS
Formulation: N-Hydroxysulfosuccinimide; powder solid
Sufficient For: 461mL of typical 1X (5mM) reaction solution, or approx. 250 uses at 2mg reagent per use
500mg Product Instructions for product #24510 Sulfo-NHS MSDS for product #24510 Sulfo-NHS Certificate of Analysis for product #24510 Sulfo-NHS $157.00

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24510B Sulfo-NHS
Formulation: N-Hydroxysulfosuccinimide; powder solid
Custom MSDS for product #24510B Sulfo-NHS Certificate of Analysis for product #24510B Sulfo-NHS
24520 Sulfo-NHS, No-Weigh Format
Formulation: N-Hydroxysulfosuccinimide; solid
Sufficient For: 15mL of typical 1X (5mM) reaction solution (total), or 8 separate uses
8 x 2mg Product Instructions for product #24520 Sulfo-NHS, No-Weigh Format MSDS for product #24520 Sulfo-NHS, No-Weigh Format Certificate of Analysis for product #24520 Sulfo-NHS, No-Weigh Format $120.00

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24525 Sulfo-NHS
Formulation: N-Hydroxysulfosuccinimide; powder solid
Sufficient For: 4605mL of typical 1X (5mM) reaction solution, or approx. 2500 uses at 2mg reagent per use
5g Product Instructions for product #24525 Sulfo-NHS MSDS for product #24525 Sulfo-NHS Certificate of Analysis for product #24525 Sulfo-NHS $1,295.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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