Iodoacetyl-LC-Biotin

碘乙酰基-LC-生物素是一种生物素化的亲电子探针，可用于研究蛋白质与亚细胞蛋白质组共价结合的范围和特征。

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货号: ajci8178
CAS: 93285-75-7
别名: N-碘代乙酰基-N'-生物素基-1,6-己二胺,N-Biotinyl-N'-(iodoacetyl)-1,6-hexanediamine,Iaa-biotin
分子式: C18H31IN4O3S
分子量: 510.43
纯度: >98%
溶解度: ≥ 51mg/mL in DMSO with gentle warming
储存: Store at -20°C
库存: 现货

Background

Iodoacetyl-LC-Biotin (N-iodoacetyl-N-biotinylhexylenediamine), a sulfhydryl-reactive biotinylation agent, is a water-insoluble reagent that requires the dissolution of suitable solvents, including dimethyl sulfoxide (DMSO) and dimethylformamide (DMF), prior to the addition into aqueous reactions. Iodoacetyl-LC-Biotin consists of a bicyclic biotin rings structure, a 1,6-diaminohexane spacer group attached to the valeric acid side chain of biotin and an iodoacetyl group at the end of the spacer providing sulfhydryl reactivity. The iodoacetyl group at the end of Iodoacetyl-LC-Biotin is able to react with sulfhydryl groups on proteins and other molecules forming a nonreversible bond. The long spacer arm of Iodoacetyl-LC-Biotin enables the modified molecule to better bind to the avidin or streptavidin probes.

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	Immunoglobulin G
Preparation method	Soluble in DMSO or DMF.
Reaction Conditions	4mM, room temperature for 90min
Applications	Immunoglobulin G (26.7 nmol) was dissolved in 1ml of 0.1M sodium phosphate, 5Mm EDTA buffer, pH 6. To this solution, DTT was added to produce a 50mM final concentration and incubation for 90min at 37 °C, the solution was allowed to cool to room temperature. Excess DTT was removed by molecular sieve chromatography using 50mM Tris, pH 8.3, with 5mM EDTA as the elution buffer. The IgG with reduced sulfhydryl groups was then biotinylated; 30 μl of 4mM iodoacetyl-LC-biotin in Me2SO was added and the mixture was incubated for 90min at room temperature in the dark. Finally, residual biotinylating reagent and free biotin were removed by dialysis against 0.1mM PBS, pH 7.2, for 72h. Protein concentrations were determined by BCA protein assay. Total biotin covalently bound to IgG was determined by an avidin-binding assay.
参考文献: [1]. Anna Bogusiewicz,Nell I. Mock, and Donald M. Mock. Instability of the biotin–protein bond in human plasma. Analytical Biochemistry 327 (2004) 156–161.