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Sulfo-NHS-LC-Biotin

In stock
Catalog No.
A8003
Amine-reactive biotinylation reagent, long chain
Grouped product items
SizePriceStock Qty
10mg
$100.00
In stock
100mg
$150.00
In stock
1g
$800.00
In stock

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Background

Sulfo-NHS-LC-biotin (Sulfosuccinimidyl 6-(biotinamido) Hexanoate) is an amine-reactive biotinylation agent. As a water-soluble analog of NHS-LC-biotin, Sulfo-NHS-LC-biotin contains a negatively charged sulfonate group on the NHS ring structure, which creates sufficient polarity within the molecule and allows it to be directly added into aqueous reactions without prior dissolution of organic solvents. Sulfo-NHS-LC-biotin covalently binds to amine groups in proteins and other molecules through its NHS ester forming an amide linkage and releasing N-Hydroxysuccinimide. Sulfo-NHS-LC-biotin has a greater length (22.4 A) between a covalently modified molecule and the bicyclic biotin rings than NHS-biotin (13.5 A), which provides the probes a greater opportunity to reach the binding sites deeply buried inside the surface plane of proteins.

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 to facilitate immobilization, purification or detection using streptavidin resins or probes
• Cell surface labeling—do not penetrate the plasma membrane, biotinylates only surface proteins of whole cells
• Amine-reactive—reacts with primary amines (-NH2), such as lysine side-chains, or the N-terminal-amine
• 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 is 22.4 angstroms; provides excellent balance between adequate length (minimal steric hindrance for biotin binding) and modest mass

Chemical Properties

StorageStore at -20°C
The product is not stable in solution, please dissolve it immediately before use.
M.Wt556.6
Cas No.127062-22-0
FormulaC20H29N4NaO9S2
SynonymsBiotinamidohexanoic acid 3-sulfo-N-hydroxysuccinimide ester sodium salt
Solubility>27.8mg/mL in DMSO
Chemical Name2,5-dioxo-1-[6-[5-(2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl)pentanoylamino]hexanoyloxy]pyrrolidine-3-sulfonic acid;sodium
SDFDownload SDF
Canonical SMILESC1C(C(=O)N(C1=O)OC(=O)CCCCCNC(=O)CCCCC2C3C(CS2)NC(=O)N3)S(=O)(=O)O.[Na]
Shipping ConditionEvaluation sample solution : ship with blue ice.All other available size: ship with RT , or blue ice upon request
General tipsFor obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.

Protocol

Biotinylation method [1]:

Sample

Cardiomyocytes

Preparation method

Soluble in Water, DMSO or DMF.

Reaction Conditions

0.5 mg/ml, 37ºC for 2 h

Applications

Sulfo-NHS-LC-Biotin was dissolved in PBS at a concentration of 0.5 mg/ml. then it was incubated with cardiomyocytes at 37ºC for 2 h. After that the cells were washed twice in cold PBS with 100 mM glycine and twice in cold PBS alone to remove the excess biotin reagent. To capture biotinylated proteins, 100 μl of streptavidin agarose resin were incubated with each 100 μg of samples for 16 hours at 4ºC. Captured proteins were eluted at 60ºC for 30 min with Laemmli 2X sample buffer which contain 40 mg/mL of dithiothreitol and they were analyzed by Western blot with primary antibodies.

References:

[1]. Charlotte Lorin, Isabelle Vögeli, Ernst Niggli. Dystrophic cardiomyopathy - role of TRPV2 channels in stretch-induced cell damage. Cardiovascular Research, 2015.

Biological Activity

Description Sulfo-NHS-LC-Biotin is an intermediate length, water-soluble biotinyltation reagent used to attach biotin to primary amines.
Targets            
IC50            

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