Biotin-azide
(Synonyms: 生物素-叠氮,N-(3-Azidopropyl)biotinamide) 目录号 : GC12570Biotin-azide叠氮生物素是一种具有叠氮基团的生物素衍生物,广泛用于生物分子标记和探针设计。
Cas No.:908007-17-0
Sample solution is provided at 25 µL, 10mM.
Biotin-azide is a biotin derivative with an azide group, widely used for biomolecule labeling and probe design[1]. Due to its azide group, Biotin-azide can undergo an azide-alkyne click chemistry reaction with molecules containing an alkyne group, forming a covalent bond. The speed, specificity, and mild reaction conditions of this reaction make it a powerful tool for biolabeling, especially for labeling biomolecules like proteins, DNA, and RNA[2-4].
In experiments, Biotin-azide can be used to link biotin to target molecules, facilitating subsequent capture, detection, and purification[5-7]. Biotin–azide probe-mediated click chemical functionalization after crosslinking enabled the isolation of transcriptional protein complexes from yeast cells[8].
References:
[1]. Goodge K, Frey M. Biotin-Conjugated Cellulose Nanofibers Prepared via Copper-Catalyzed Alkyne-Azide Cycloaddition (CuAAC) "Click" Chemistry. Nanomaterials (Basel). 2020 Jun 16;10(6):1172. doi: 10.3390/nano10061172. PMID: 32560117; PMCID: PMC7353318.
[2]. Tsuji T, Fujimoto T. Ultrastructural localization of de novo synthesized phosphatidylcholine in yeast cells by freeze-fracture electron microscopy. STAR Protoc. 2021 Dec 6;2(4):100990. doi: 10.1016/j.xpro.2021.100990. PMID: 34934959; PMCID: PMC8654986.
[3]. Guo T, Zhou L, et,al. N-homocysteinylation of DJ-1 promotes neurodegeneration in Parkinson's disease. Aging Cell. 2024 May;23(5):e14124. doi: 10.1111/acel.14124. Epub 2024 Feb 21. PMID: 38380563; PMCID: PMC11113254.
[4]. Bruckman MA, Czapar AE, et,al. Tobacco mosaic virus-based protein nanoparticles and nanorods for chemotherapy delivery targeting breast cancer. J Control Release. 2016 Jun 10;231:103-13. doi: 10.1016/j.jconrel.2016.02.045. Epub 2016 Mar 3. PMID: 26941034; PMCID: PMC5207211.
[5]. Zhou L, Guo T, et,al. N-homocysteinylation of α-synuclein promotes its aggregation and neurotoxicity. Aging Cell. 2023 Mar;22(3):e13745. doi: 10.1111/acel.13745. Epub 2022 Nov 27. PMID: 36437524; PMCID: PMC10014048.
[6]. Bruckman MA, Czapar AE, et,al. Tobacco mosaic virus-based protein nanoparticles and nanorods for chemotherapy delivery targeting breast cancer. J Control Release. 2016 Jun 10;231:103-13. doi: 10.1016/j.jconrel.2016.02.045. Epub 2016 Mar 3. PMID: 26941034; PMCID: PMC5207211.
[7]. Kim HY, Tallman KA, et,al. An azido-biotin reagent for use in the isolation of protein adducts of lipid-derived electrophiles by streptavidin catch and photorelease. Mol Cell Proteomics. 2009 Sep;8(9):2080-9. doi: 10.1074/mcp.M900121-MCP200. Epub 2009 May 30. PMID: 19483245; PMCID: PMC2742437.
[8]. Zheng S, Lin J, et,al. Aberrant Cholesterol Metabolism and Wnt/β-Catenin Signaling Coalesce via Frizzled5 in Supporting Cancer Growth. Adv Sci (Weinh). 2022 Oct;9(28):e2200750. doi: 10.1002/advs.202200750. Epub 2022 Aug 17. PMID: 35975457; PMCID: PMC9534957.
Biotin-azide叠氮生物素是一种具有叠氮基团的生物素衍生物,广泛用于生物分子标记和探针设计。[1]Biotin-azide由于其叠氮化物基团的存在,可以与含有炔基的分子发生叠氮化物-炔键化学反应,形成共价键。该反应的速度、特异性和温和的反应条件使其成为生物标记的有力工具,特别是用于标记蛋白质、DNA和RNA等生物分子[2-4]。
在实验中,Biotin-azide可以用来连接生物素和目标分子,促进随后的捕获、检测和纯化[5-7],例如:Biotin-azide探针交联后的点击化学功能化使得从酵母细胞中分离转录蛋白复合物成为可能[8]。
一、Biotin-azide点击化学和下拉实验[1]:
1. 用50µM棕榈酸类似物Alk-16处理细胞5h,收集细胞并在加入protease inhibitor cocktail 的1% NP-40裂解缓冲液(25mM Tris-HCl pH 8.0, 150mM NaCl, 10% glycerol, 1% NP-40)中裂解。
2. 在4℃下14000rpm离心20min后收集上清。BCA测定蛋白浓度。
3. 按以下顺序加入化学试剂: 1µL 10mM Biotin-azide, 1.2µL 10mM tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA), 1µL 40mM CuSO4,1µL 40 mM tris(2-carboxyethyl)phosphine HCl (TCEP hydrochloride) 。
4. 将反应混合物充分混合,在室温下黑暗孵育30min,加入10µL 0.5M EDTA使反应猝灭,并短暂旋转样品。
5. 将pellet重悬于50µL TEA缓冲液中,并在室温下旋转10min。一旦pellet重悬,加入450µL TEA缓冲液并短暂旋转。
6. 用TEA缓冲液洗涤15µL Pierce Streptavidin琼脂糖3次,加入每个样品的上清液中,在室温下旋转孵育90min。
7. 用1mL 1% SDS在PBS中洗涤beads三次。每个样品在50µL裂解缓冲液中重悬,然后在95℃下加热5min。
8. 取15µL样品进行Western Blot分析。
本程序仅提供指导,请根据您的具体需求进行修改。
References:
[1]. Zheng S, Lin J, et,al. Aberrant Cholesterol Metabolism and Wnt/β-Catenin Signaling Coalesce via Frizzled5 in Supporting Cancer Growth. Adv Sci (Weinh). 2022 Oct;9(28):e2200750. doi: 10.1002/advs.202200750. Epub 2022 Aug 17. PMID: 35975457; PMCID: PMC9534957.
Cas No. | 908007-17-0 | SDF | |
别名 | 生物素-叠氮,N-(3-Azidopropyl)biotinamide | ||
化学名 | N-(3-azidopropyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide | ||
Canonical SMILES | O=C1N[C@@]([H])(CS[C@H]2CCCCC(NCCCN=[N+]=[N-])=O)[C@]2([H])N1 | ||
分子式 | C13H22N6O2S | 分子量 | 326.42 |
溶解度 | ≥ 32.6mg/mL in DMSO, ≥ 2.51 mg/mL in EtOH with ultrasonic | 储存条件 | Store at -20°C |
General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 储备液的保存方式和期限:-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。 为了提高溶解度,请将管子加热至37℃,然后在超声波浴中震荡一段时间。 |
||
Shipping Condition | 评估样品解决方案:配备蓝冰进行发货。所有其他可用尺寸:配备RT,或根据请求配备蓝冰。 |
制备储备液 | |||
1 mg | 5 mg | 10 mg | |
1 mM | 3.0635 mL | 15.3177 mL | 30.6354 mL |
5 mM | 0.6127 mL | 3.0635 mL | 6.1271 mL |
10 mM | 0.3064 mL | 1.5318 mL | 3.0635 mL |
第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量) | ||||||||||
给药剂量 | mg/kg | 动物平均体重 | g | 每只动物给药体积 | ul | 动物数量 | 只 | |||
第二步:请输入动物体内配方组成(配方适用于不溶于水的药物;不同批次药物配方比例不同,请联系GLPBIO为您提供正确的澄清溶液配方) | ||||||||||
% DMSO % % Tween 80 % saline | ||||||||||
计算重置 |
计算结果:
工作液浓度: mg/ml;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL,
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL saline,混匀澄清。
1. 首先保证母液是澄清的;
2.
一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet