NHS-LC-Biotin

Biotinylation reagent

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100mg

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货号: ajci9408
CAS: 72040-63-2
别名: 生物素化-epsilon-氨基己酸-N-羟基丁二酰亚胺活化酯,Succinimidyl 6-(biotinamido)hexanoate
分子式: C20H30N4O6S
分子量: 454.54
纯度: >98%
溶解度: ≥ 22.727mg/mL in DMSO
储存: Store at -20°C, protect from light, unstable in solution, ready to use.
库存: 现货

Background

NHS-LC-biotin (succinimidyl-6-(biotinamindo)hexanoate), also known as NHS-X-biotin is a derivative of _D-biotin, a amine-reactive biotinylation agent, that contains a spacer arm off the valeric acid side chain of _D-biotin with an NHS ester group at its end. The NHS ester group at the end of NHS-LC-biotin covalently binds to amine groups in proteins and other molecules forming a stable amide linkage and releasing the NHS group. The 6-aminocaproic acid spacer of NHS-LC-biotin greatly increases the length between a covalently modified molecule and the bicyclic biotin rings leading to a better binding potential for avidin or streptavidin probes. NHS-LC-biotin is insoluble in aqueous environments requiring the dissolution of organic solvents prior to the addition to a buffered reaction.

Reference

[1].Bioconjugate Techniques , 2nd ed. By Greg T.Hermanson ?(Pierce Biotechnology, Thermo Fisher Scientific, Rockford, IL). ?Academic Press ?(an imprint of Elsevier): ?London, Amsterdam, Burlington, San Diego . 2008. ISBN 978-0-12-370501-3.

Protocol

Biotinylation method [1]:
Sample	S. aureus
Preparation method	Soluble in DMSO or DMF.
Reaction Conditions	60 mg/ml, room temperature for 1 h.
Applications	Fixed cell suspensions of S. aureus were diluted to a 2% cell suspension in 0.2 M NaHCO3, pH 8.3. NHS-LC-biotin freshly dissolved in DMSO at a concentration of 60 mg/ml, was added to the cell suspension subsequently. After gentle tumble mixing for 60min at room temperature, the reaction was stopped by removal of excess biotin via gel ?ltration over a PD-10 column and washed extensively in 0.01 mM Tris-buffered 0.15 M NaCl (TBS). Successful protein biotinylation was examining their reactivity with alkaline phosphatase-conjugated streptavidin（AP-STRAV). AP-STRAV binding was detected as a function of pNPP hydrolysis.
参考文献: [1]. TRUC NGUYEN, BERHANE GHEBREHIWET, AND ELLINOR I. B. PEERSCHKE. Staphylococcus aureus Protein A Recognizes Platelet gC1qR/p33: a Novel Mechanism for Staphylococcal Interactions with Platelets. INFECTION AND IMMUNITY 2000, p. 2061–2068.