EZ-Link Sulfo-NHS-LC-LC-Biotin

Thermo Scientific EZ-Link Sulfo-NHS-LC-LC-Biotin is a long-chain biotinylation reagent for labeling antibodies, proteins and other molecules that have primary amines.

Sulfo-NHS-LC-LC-Biotin is the longest 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.

Highlights:

• 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
• Doubly long – spacer arm (total length added to target) is 22.4 angstroms; this extended arm helps to minimize steric hindrance for biotin binding

Product Details:

Chemical structure of Sulfo-NHS-LC-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.

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.

References:

1. Daniels, G.M. and Amara, S.G. (1998). Selective labeling of neurotransmitter transporters at the cell surface. Methods Enzymol. 296:307-18.
2. Huh, K-H. and Wenthold, R.J. (1999). Turnover analysis of glutamate receptors identifies a rapidly degraded pool of the N-methyl-D-aspartate receptor subunit, NR1, in cultured cerebellar granule cells. J. Biol. Chem. 274:151-7.
3. Ali, M.K. and Bergson, C. (2003). Elevated intracellular calcium triggers recruitment of the receptor cross-talk accessory protein calcyon to the plasma membrane. J. Biol. Chem. 278(51): 51654-63.
4. Borroto, A., et al. (2003). Impaired trafficking and activation of tumor necrosis factor-alpha-converting enzyme in cell mutants defective in protein ectodomain shedding. J. Biol. Chem. 278(28):
   25933-9.
5. Chyung, J.H. and Selkoe, D.J. (2003). Inhibition of receptor-mediated endocytosis demonstrates generation of amyloid protein at the cell surface. J. Biol. Chem. 278(51): 51035-43.
6. Frickel, E-M., et al. (2002). TROSY-NMR reveals interaction between Erp57 and the tip of the calreticulin P-domain. PNAS 99(4): 1954-9.
7. Gimferrer, I. , et al. (2003). The accessory molecules CD5 and CD6 associate on the membrane of lymphoid T cells. J. Biol. Chem. 278(10): 8564-71.
8. Li, H. and Pajor, A.M. (2003). Mutagenesis of the N-glycosylation site of hNaSi-1 reduces transport activity. Am. J. Physiol. Cell Physiol. 285:C1188-96.
9. Lukashevich, I.S., et al. (2003). Arenavirus-mediated liver pathology: acute lymphocytic choriomeningitis virus infection of rhesus macaques is characterized by high-level interleukin-6 expression and hepatocyte proliferation. J. Virol. 77(3): 1727-37.
10. Ohnishi, T., et al. (2003). MD-2 is necessary for the toll-like receptor 4 protein to undergo glycosylation essential for its translocation to the cell surface. Clin. Diagn. Lab. Immunol. 10(3): 405-410.

Related Resources:

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

Related Products:

Sulfo-NHS-Biotin and Kits – 13.5 angstrom spacer arm
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Sulfo-NHS-SS-Biotin and Kits – 24.3 angstrom spacer arm, cleavable
All Amine-Reactive Biotinylation Reagents
All Biotinylation Reagents
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