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EZ-Link™ Sulfo-NHS-LC-Biotin
EZ-Link™ Sulfo-NHS-LC-Biotin
EZ-Link™ Sulfo-NHS-LC-Biotin
EZ-Link™ Sulfo-NHS-LC-Biotin
EZ-Link™ Sulfo-NHS-LC-Biotin
Thermo Scientific™

EZ-Link™ Sulfo-NHS-LC-Biotin

Thermo Scientific EZ-Link Sulfo-NHS-LC-Biotin is an intermediate-length, water-soluble biotinylation reagent for labeling antibodies, proteins and other molecules that have primaryRead more
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Catalog NumberQuantity
21335100 mg
A3925710 x 1 mg
Catalog number 21335
Price (USD)
389.00
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100 mg
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Thermo Scientific EZ-Link Sulfo-NHS-LC-Biotin is an intermediate-length, water-soluble biotinylation reagent for labeling antibodies, proteins and other molecules that have primary amines.

Features of EZ-Link Sulfo-NHS-LC-Biotin:

Protein labeling—biotinylate antibodies to facilitate immobilization, purification or detection using streptavidin resins or probes
Cell surface labeling—biotinylates only surface proteins of whole cells because the negatively charged reagent does not permeate cell membranes
Amine-reactive—reacts with primary amines (-NH2), such as lysine side-chains, or the amino-termini of polypeptides
Soluble—charged sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds
Irreversible—forms permanent amide bonds; spacer arm cannot be cleaved
Medium length—spacer arm (total length added to target) is 22.4 angstroms; provides excellent balance between adequate length (minimal steric hindrance for biotin binding) and modest mass

Sulfo-NHS-LC-Biotin is one of three very similar EZ-Link Reagents that are water-soluble, non-cleavable, and enable simple and efficient biotinylation of antibodies, proteins and any other primary amine-containing macromolecules in solution. Specific labeling of cell surface proteins is another common application for these uniquely water-soluble and membrane impermeable reagents. Differing only in their spacer arm lengths, the three Sulfo-NHS-ester reagents offer the possibility of optimizing labeling and detection experiments where steric hindrance of biotin binding is an important factor. Sulfo-NHS-LC-Biotin is offered in several package sizes and as complete protein labeling kits.

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.

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Pierce™ Premium Grade Sulfo-NHS-LC-Biotin
WARNING: Reproductive Harm - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cell PermeabilityCell-Impermeant
DescriptionEZ-Link Sulfo-NHS-LC-Biotin
Label TypeBiotin & Analogs
Product LineEZ-Link™
Product TypeSulfo-NHS-LC-Biotin
Quantity100 mg
Reactive MoietyActive Ester, Succinimidyl Ester, Sulfo-NHS Ester
Chemical ReactivityAmine
Label or DyeBiotin
SolubilityDMF (Dimethylformamide), DMSO (Dimethylsulfoxide), Water
SpacerMid-length
Unit SizeEach
Contents & Storage
Store desiccated at -20°C. Shipped at ambient temperature.
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Certificates

Lot #Certificate TypeDateCatalog Number(s)
3215142Certificate of AnalysisJun 25, 2025A39257
3213819Certificate of AnalysisJun 18, 202521335
3214540Certificate of AnalysisJun 02, 2025A39257
3212137Certificate of AnalysisMay 20, 202521335
3212453ACertificate of AnalysisMay 05, 2025A39257
5 results displayed, search above for a specific certificate

Safety Data Sheets

Frequently asked questions (FAQs)

No. NHS esters hydrolyze very quickly. The container must be brought to room temperature (RT) prior to opening, then the amount of biotinylation reagent weighed, dissolved and used immediately.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Yes. Because this molecule dissolves readily in polar solutions and is itself charged by the sodium sulfoxide group on the succinimidyl ring, it cannot permeate the cell wall or membrane of an intact cell. As long as the cell is intact, only the primary amines on the surface will be biotinylated.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Tris is a primary amine, which will directly compete with protein primary amines for reaction with the NHS-ester activated biotin reagent. However, once a protein is labeled and desalted, Tris buffer can be used for storage or assay buffers, as appropriate for the protein and application.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

The molar excess to use depends on the concentration of the ligand to be biotinylated. Although the literature contains references that use between 2-50 molar excess of biotin, we recommend the following: 10-fold molar excess for 10 mg/mL solution of protein; 25-fold molar excess for 2 mg/mL solution of protein.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

No, the EZ-Link Sulfo-NHS-LC-Biotin, No-Weigh Format (Cat. No. A39257) should not be dissolved in water for storage. We recommended storing EZ-Link Sulfo-NHS-LC-Biotin, No-Weigh Format at -20 degrees C with desiccant as it is very moisture sensitive. Furthermore, to avoid moisture condensation onto the product, equilibrate vial to room temperature before opening.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Citations & References (22)

Citations & References
Abstract
Rapid identification of highly potent human anti-GPCR antagonist monoclonal antibodies.
Authors:Scott MJ, Jowett A, Orecchia M, Ertl P, Ouro-Gnao L, Ticehurst J, Gower D, Yates J, Poulton K, Harris C, Mullin MJ, Smith KJ, Lewis AP, Barton N, Washburn ML, de Wildt R
Journal:
PubMed ID:32343620
'Complex cellular targets such as G protein-coupled receptors (GPCRs), ion channels, and other multi-transmembrane proteins represent a significant challenge for therapeutic antibody discovery, primarily because of poor stability of the target protein upon extraction from cell membranes. To assess whether a limited set of membrane-bound antigen formats could be exploited ... More
Potent neutralizing monoclonal antibodies against Ebola virus isolated from vaccinated donors.
Authors:Fan P, Chi X, Liu G, Zhang G, Chen Z, Liu Y, Fang T, Li J, Banadyga L, He S, Yu C, Qiu X, Chen W
Journal:
PubMed ID:32213108
Ebola virus (EBOV) can cause severe hemorrhagic fever in humans, and no approved treatment is currently available. Although several antibodies have achieved good protection in animal models, the potential emerging isolates of ebolavirus and the unknown effects of experimental antibodies in humans underscore the need to develop additional antibodies to ... More
Junctional and somatic hypermutation-induced CX
Authors:Yi C, Xia J, He L, Ling Z, Wang X, Yan Y, Wang J, Zhao X, Fan W, Sun X, Zhang R, Ye S, Zhang R, Xu Y, Ma L, Zhang Y, Zhou H, Huang Z, Niu J, Long G, Lu J, Zhong J, Sun B
Journal:Cell Mol Immunol
PubMed ID:32235917
'Induction of broadly neutralizing monoclonal antibodies (bNAbs) that bind to the viral envelope glycoproteins is a major goal of hepatitis C virus (HCV) vaccine research. The study of bNAbs arising in natural infection is essential in this endeavor. We generated a human antibody, 8D6, recognizing the E2 protein of HCV ... More
Mechanisms of tethering and cargo transfer during epididymosome-sperm interactions.
Authors:Zhou W, Stanger SJ, Anderson AL, Bernstein IR, De Iuliis GN, McCluskey A, McLaughlin EA, Dun MD, Nixon B
Journal:BMC Biol
PubMed ID:30999907
'The mammalian epididymis is responsible for the provision of a highly specialized environment in which spermatozoa acquire functional maturity and are subsequently stored in preparation for ejaculation. Making important contributions to both processes are epididymosomes, small extracellular vesicles released from the epididymal soma via an apocrine secretory pathway. While considerable ... More
Molecular determinants in Frizzled, Reck, and Wnt7a for ligand-specific signaling in neurovascular development.
Authors:Cho C, Wang Y, Smallwood PM, Williams J, Nathans J
Journal:Elife
PubMed ID:31225798
'The molecular basis of Wnt-Frizzled specificity is a central question in developmental biology. Reck, a multi-domain and multi-functional glycosylphosphatidylinositol-anchored protein, specifically enhances beta-catenin signaling by Wnt7a and Wnt7b in cooperation with the 7-transmembrane protein Gpr124. Among amino acids that distinguish Wnt7a and Wnt7b from other Wnts, two clusters are essential ... More
22 total citations

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