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EZ-Link NHS-LC-Biotin

Long-chain, yet simple and membrane-permeable, amine-reactive biotinylation reagent.

EZ-Link NHS-LC-Biotin

Thermo Scientific EZ-Link NHS-LC-Biotin is a long-chain, NHS-ester activated biotinylation reagent for labeling primary amines (e.g., protein lysines), whose membrane permeability enables it to be used for general intracellular labeling.

NHS-LC-Biotin is succinimidyl-6-(biotinamido)hexanoate. It is one of three similar EZ-Link NHS-Biotin Reagents that enable simple and efficient biotinylation of antibodies, proteins and any other primary amine-containing biomolecules in solution. Differing only in their spacer arm lengths, the three NHS-ester reagents offer researchers the possibility of optimizing labeling and detection experiments where steric hindrance of biotin binding is an important factor. Because they are uncharged and contain simple alkyl-chain spacer arms, these biotin compounds are membrane-permeable and useful for intracellular labeling.


  • Protein labeling – biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins
  • Membrane-permeable – can be used to label inside cells (intracellular)
  • Amine-reactive – reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the amino-termini of polypeptides
  • Irreversible – forms permanent amide bonds; spacer arm cannot be cleaved
  • Solubility – must be dissolved in DMSO or DMF before further dilution in aqueous buffers
  • Medium length – spacer arm (total length added to target) is 22.4 angstroms; it consists of the biotin valeric acid group extended by a 6-atom chain

Product Details:

Chemical structure of NHS-LC-Biotin Chemical structure of NHS-LC-Biotin. This biotinylation reagent labels antibodies and other proteins and molecules that contain primary amines (e.g., side-chain of lysine). For more information, see our review of NHS Ester Reaction Chemistry.

Properties of EZ-Link NHS-Biotin Reagents. These compounds have simple alkyl spacer arms and are cell membrane permeable. Reagents with Sulfo-NHS groups and/or pegylated or cleavable spacer arms are also available.
  NHS-Biotin NHS-LC-Biotin NHS-LC-LC-Biotin
Molecular formula C14H19O5N3S C20H30O6N4S C26H41O7N5S
Molecular weight 341.38 454.54 567.70
Spacer arm length 13.5 angstroms 22.4 angstroms 30.5 angstroms
Mass added to target 226.3 daltons 339.5 daltons 452.6 daltons
CAS number 35013-72-0 72040-63-2  
Form White powder White to off-white powder White to off-white powder
SMILES structures O=C(CC1)N(OC(CCCCC(SC2)C3C2NC(N3)=O)=O)C1=O
Storage conditions 4°C with desiccant, protect from moisture, use only fresh solutions
Reactive groups N-hydroxysuccinimide ester, reacts with primary amines at pH 7.0-9.0

We manufacture biotin reagents to ensure the highest possible overall product integrity, consistency and performance for the intended research applications.

N-Hydroxysulfosuccinimide (NHS) esters of biotin are the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.

Varieties of biotin NHS-ester reagents differ in length, solubility, cell permeability and cleavability. Non-sulfonated NHS-biotins are cell permeable but must be dissolved in organic solvent such as DMSO or DMF. Sulfo-NHS biotins (and those with pegylated spacers) are directly water soluble but not membrane permeable. Varieties containing disulfide bonds can be cleaved using reducing agents, enabling the biotin group to be disconnected from the labeled protein.


  1. Fouassier, L., et al. (2000) J. Biol. Chem. 275 , 25039-25045.
  2. Nunomura, W., et al. (2000) J. Biol. Chem. 275 , 6360-6367.
  3. Chiu , N.H. , et al. (1999) Clin. Chem.45 , 1954-1959.
  4. Schumacher, T.N.M., et al. (1996). Science 271, 1854-1857.
  5. Burgess, T.L., et al. (1995). J. Biol. Chem.270, 19188-19196.

Related Resources:

Biotinylation Reagents Selection Guide
Review of Biotinylation Methods and Applications
Chemistry of Crosslinking (and Labeling Reagents)

Related Products:

All Amine-Reactive Biotinylation Reagents
All Biotinylation Reagents
Biotin Quantitation Reagents and Kits
Avidin, Streptavidin, NeutrAvidin Affinity Resins

Ordering Information
Product # Description Pkg. Size Instructions MSDS CofA Price
21336 EZ-Link NHS-LC-Biotin
Formulation: NHS ester of biotin with a 6-aminocaproic acid spacer
Sufficient For: Approx. 25 uses to label a total of 733mg of IgG using typical conditions
50mg Product Instructions for product #21336 EZ-Link NHS-LC-Biotin MSDS for product #21336 EZ-Link NHS-LC-Biotin Certificate of Analysis for product #21336 EZ-Link NHS-LC-Biotin $138.00

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21336B EZ-Link NHS-LC-Biotin
Formulation: NHS ester of biotin with a 6-aminocaproic acid spacer
Custom MSDS for product #21336B EZ-Link NHS-LC-Biotin Certificate of Analysis for product #21336B EZ-Link NHS-LC-Biotin

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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