Thermo Scientific EZ-Link Sulfo-NHS-Biotin is a short-chain, water-soluble biotinylation reagent for labeling antibodies, proteins and other molecules that have primary amines.
Sulfo-NHS-Biotin is the shortest 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-Biotin is offered in several package sizes and as complete protein labeling kits.
- 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 the side-chain of lysines (K) 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
- Very short – spacer arm (total length added to target) is 13.5 angstroms; it consists of the native biotin valeric acid group only, adding the small possible
||Chemical structure of Sulfo-NHS-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 Sulfo-NHS-Biotin Labeling Reagents. These compounds have simple alkyl spacer arms and water-soluble Sulfo-NHS groups. Reagents with non-sulfonated NHS-ester groups (membrane-permeable) and/or pegylated or cleavable spacer arms are also available.
|Spacer arm length
|Mass added to target
||White to off-white powder
||White to tan powder
||White to off-white powder
||-20°C with desiccant, protect from moisture, use only fresh solutions
||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.
- Hoffmann, A., et al. (2003). J. Neuroscience 23(11), 4410-4419.
- Dorsey, C.W., et al. (2003). Microbiology 149, 1227-1238.
- Make, J.L. and Dickerson, H.W. (2003). Clin. Diag. Lab Immunol. 10, 876-881.
- Usui, K., et al. (2004). Protein Science 13, 134-144.
Biotinylation Reagents Selection Guide
Review of Biotinylation Methods and Applications
Chemistry of Crosslinking (and Labeling Reagents)
Sulfo-NHS-LC-Biotin and Kits – 22.4 angstrom spacer arm
NHS-PEG4-Biotin and Kits – 29.0 angstrom spacer arm, pegylated
Sulfo-NHS-SS-Biotin and Kits – 24.3 angstrom spacer arm, cleavable
All Amine-Reactive Biotinylation Reagents
All Biotinylation Reagents
Biotin Quantitation Reagents and Kits
Avidin, Streptavidin, NeutrAvidin Affinity Resins