JavaScript seems to be disabled in your browser. For the best experience on our site, be sure to turn on Javascript in your browser.
Tel: +1-832-696-8203
Email: [email protected]
Worldwide Distributors
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Sulfo-NHS-Biotin (N-Hydroxysulfosuccinimidobiotin) is an amine-reactive biotinylation reagent. The most common amine-reactive biotinylation reagents can be divied into two basic types, N-hydroxysuccinimide (NHS) esters and carboxylates. Generally, NHS esters biotinylation reagent contains a reactive NHS ring structure that reacts with an amine on the carbonyl group of a protein or other molecules through nucleophilic attack subsequently forming a stable amide linkage and releasing the NHS group. Due to the insolubility of NHS-biotin in aqueous environments, sulfo-NHS-biotin, a water-soluble analog of NHS-biotin, has been developed by attaching a negatively charged sulfonate group on the NHS ring structure, which can be added directly to aqueous reactions without the need for organic solvent dissolution.
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.
• 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 • Solubility—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 is 13.5 angstroms; it consists of the native biotin valeric acid group only
Sample
THUMPa protein
Preparation method
Soluble in water, DMSO or DMF.
Reaction Conditions
2 mM, room temperature for 30 min
Applications
Sulfo-NHS-biotin reagents were dissolved at 2 mM in a 50-mM K2HPO4/KH2PO4 buffer (pH 7.5) containing 50 mM of NaCl. They were immediately used in separated assays to modify 1.66 nmol of THUMPa protein. The labeling reactions were incubated 30 min at room temperature. The samples were then dialyzed for 15 min against a 50-mM K2HPO4/KH2PO4 buffer (pH 7.5) containing 50 mM of NaCl. TTotal biotin covalently bound to proteins was determined by an avidin-binding assay.
References:
[1]. Guillaume Gabant, Julie Augier and Jean Armengaud. Assessment of solvent residues accessibility using three Sulfo-NHS-biotin reagents in parallel: application to footprint changes of a methyltransferase upon binding its substrate. J. Mass Spectrom. 2008; 43: 360–370.
