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NHS-LC-Biotin

NHS-LC-Biotin

Catalog No.

A8004

Amine-reactive biotinylation reagent, long chain

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Size	Price	Stock	Qty
100mg	$60.00	In stock

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Background
Product Citation
Features
Chemical Properties
Protocol
Biological Activity
Quality Control

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

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.

Product Citation

Features

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

Chemical Properties

Storage	Store at -20°C
M.Wt	454.54
Cas No.	72040-63-2
Formula	C₂₀H₃₀N₄O₆S
Synonyms	Succinimidyl 6-(biotinamido)hexanoate
Solubility	≥22.73 mg/mL in DMSO; insoluble in EtOH; insoluble in H2O
Chemical Name	(2,5-dioxopyrrolidin-1-yl) 6-[5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]hexanoate
SDF	Download SDF
Canonical SMILES	C1CC(=O)N(C1=O)OC(=O)CCCCCNC(=O)CCCCC2C3C(CS2)NC(=O)N3
Shipping Condition	Ship with blue ice, or upon other requests.
General tips	For obtaining a higher solubility, please warm the tube at 37°C and shake it in the ultrasonic bath for a while. We do not recommend long-term storage for the solution, please use it up soon.

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 ﬁltration 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.
References: [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.