|EZLink Sulfo-NHS-Biotin Kit
Amine-reactive biotinyltation kit, water-soluble
Sample solution is provided at 25 µL, 10mM.
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Components and storage
|Sufficient For: 10 labeling reactions, each with 1 to 10 mg of antibody|
|Streptavidin||10 mg||EZ-Link Sulfo-NHS-Biotin||25 mg|
|HABA Solution||1 mL||PBS Pack (makes 500 mL)||1 pack|
|Sephadex G-25 in PD-10 Desalting Columns||8.3 mL, 10 columns|
|Store biotin and streptavidin reagents at -20°C. Store remaining kit components at 4°C.|
|Label or Dye:||Biotin||Product Size:||10 reactions|
|Labeling Scale:||1-10 mg||Labeling Target:||Proteins (General), Antibodies (General)|
Clearance of components and concentrations.
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• Amine-reaction—Reacting with primary amines (-NH2), such as lysine side-chains, or the amino-termini of polypeptides.
• Labeling antibody—This kit can label antibodies to facilitate immobilization, purification or detection.
• Labeling protein—This kit can label proteins to facilitate immobilization, purification or detection.
• Labeling Cell surface molecule—This kit can label the cell surface proteins because the negatively charged reagent does not permeate cell membranes.
• Irreversible-Permanent amide bonds are formed; Spacer arm cannot be cleaved.
• Very short arm—Spacer arm (total length added to target) is 13.5 angstroms; it consists of the native biotin valeric acid group only.
• Solubility increased—Sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds.
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.