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NHS-SS-Biotin

现货
Catalog No.
A8006
生物素化试剂
组合的产品项目
规格价格库存 数量
100mg
¥ 1,000.00
现货
1g
¥ 7,000.00
现货

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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.

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.

Features

• Protein labeling—biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins
• Intracellular labeling—membrane-permeable, can be used to label inside cells
• Amine-reactive—reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the N-terminal-amine
• Cleavable—disulfide bond in spacer arm allows the biotin label to be removed using reducing agents such as DTT
• Solubility—water insoluble, must be dissolved in DMSO or DMF before further dilution in aqueous buffers
• Medium length—spacer arm is 24.3 angstroms; it consists of the native biotin valeric acid group extended by a 7-atom chain

Chemical Properties

StorageStore at -20°C
该产品在溶液中不稳定,建议现配现用
M.Wt504.64
FormulaC19H28N4O6S3
SynonymsNHS-SS-Biotin,Biotin disulfide N-hydroxysuccinimide ester
Solubility≥28.786mg/mL in DMSO, <1.98 mg/mL in EtOH, <1.97 mg/mL in H2O
Chemical Name2,5-dioxopyrrolidin-1-yl 3-((2-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)ethyl)disulfanyl)propanoate
SDFDownload SDF
Canonical SMILESO=C1CCC(N1OC(CCSSCCNC(CCCC[C@@H]2SC[C@@](N3)([H])[C@]2([H])NC3=O)=O)=O)=O
运输条件试用装:蓝冰运输。 其他可选规格:常温运输或根据您的要求用蓝冰运输。
一般建议为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。

试验操作

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 artificial cerebrospinal fluid(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 buffer (50 mM glutathione, 75 mM NaCl, 10 mM EDTA, 1% BSA, and 0.075 N NaOH). Neurons were lysed in the modified RIPA buffer (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.

References:

[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.

生物活性

描述 NHS-SS-Biotin是一种细胞通透性的和伯胺反应性的生物素化试剂。
靶点 proteins          
IC50            

质量控制

化学结构

NHS-SS-Biotin

相关生物数据

NHS-SS-Biotin