NHS-SS-Biotin

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NHS-SS-Biotin

本产品不向个人销售，仅用作科学研究，不用于任何人体实验及非科研性质的动物实验。

规格:	98%
分子量:	504.64

包装与价格：

包装	价格(元)
100mg	电议
1g	电议

产品介绍

NHS-SS-生物素是一种可切割的 ADC 接头，用于合成抗体-药物偶联物 (ADC)。
货号:ajcx11150
CAS:122266-55-1
分子式:C19H28N4O6S3
分子量：504.64
溶解度:≥ 28.786mg/mL in DMSO
纯度：98%
存储：Store at -20°C
库存：现货

Background:

NHS-SS-biotin (Succinimidyl-2-(biotinamido)-ethyl-1,3-dithiopropionate) is a long-chain cleavable amine-reactive biotinylation reagent that has a spacer arm of 24.3 A in length due to the existence of a cross-bridge within its chemical structure. The long spacer arm creates an increased length between a covalently modified molecule and the bicyclic biotin rings that greatly enhances the ability of the molecule to bind to avidin or streptavidin probes. The NHS ester group of NHS-SS-biotin reacts with the amine groups of proteins and other molecule forming an amide bond and releasing of sulfo-NHS. NHS-SS-Biotin is a water-insoluble molecule that requires the dissolution of organic solvents prior to be added to aqueous reactions.

参考文献

[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	hippocampal neurons
Preparation method	Soluble in DMSO or DMF.
Reaction Conditions	1.5 mg/ml, 4 ℃ for 1 h
Applications	Neurons were washed with the arti?cial cerebrospinal ?uid（ACSF） at 37 ℃, and incubated with 1.5 ml of 1.5 mg/ml NHS-SS-biotin with gentle shaking at 4 ℃ for 1 h. After washing, neurons were switched to the neuronal culture medium and incubated at 37 ℃. At indicated times of incubation, neurons were cooled to 4 ℃ and un-endocytosed surface biotin was cleaved by incubating in the glutathione cleavage bu?er (50 mM glutathione, 75 mM NaCl, 10 mM EDTA, 1% BSA, and 0.075 N NaOH). Neurons were lysed in the modi?ed RIPA bu?er (50 mM Tris–HCl, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxylate, 1 mM EDTA, and protease inhibitors). Lysates were cleared by centrifugation at 10,000g for 10 min at 4 ℃ and incubated at 4 ℃ over-night with 70 μl of 50% streptavidin beads. Endocytosis of ErbB proteins was assayed using cleavable biotin. Bead-associated proteins were subjected to Western blot analysis.
参考文献: [1]. Yu Liu, Yan-Mei Tao, Ran-Sook Woo, Wen-Cheng Xiong, Lin Mei. Stimulated ErbB4 internalization is necessary for neuregulin signaling in neurons. Biochemical and Biophysical Research Communications 354 (2007) 505–510.