客户使用产品发表文献 2 篇

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Quality Control & SDS

View current batch:

Purity: >98.00%

实验参考方法

Biotinylation method [1]:
Sample	whole blood
Preparation method	Soluble in DMSO or DMF.
Reaction Conditions	30 minutes
Applications	Firstly, NHS-biotin was dissolved in DMSO at a concentration of 100 mg/mL, after that using 90% DMSO solution diluted the biotin to a concentration of 20 mg/mL. The biotin solution was sterilized by ﬁltering the solution through 0.2-μm pore nylon syringe ﬁlters and the solution was diluted with 6 volumes 0.9% saline solution. Blood was allowed to warm to room temperature prior to labeling with NHS-biotin. To achieve a biotin concentration of 0.04 pg of biotin/RBC（Red Blood Cells）, Biotin-DMSO-saline solution was injected into each blood bag and agitated for 30 minutes.
References: [1]. J.E. Tomlinson, E. Taberner, R.C. Boston, S.D. Owens, and R.D. Nolen-Walston. Survival Time of Cross-Match Incompatible Red Blood Cells in Adult Horses. J Vet Intern Med 2015;29:1683–1688.

产品描述

NHS-Biotin (C₁₄H₁₈O₅N₃S) is N-hydroxysuccinimido biotin. It is the shortest of three similar EZ-Link NHS-Biotin Reagents that enable simple and efficient biotinylation of antibodies, proteins and any other primary amine-containing biomolecules in solution. NHS-Biotin offers researchers the possibility of optimizing labeling and detection experiments where steric hindrance of biotin binding is an important factor. Because it is uncharged and contain simple alkyl-chain spacer arms, NHS-Biotin compound is membrane-permeable and useful for intracellular labeling.

NHS-Biotin is the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.

NHS-Biotin（C14H18O5N3S）是N-羟基琥珀酰亚胺生物素。它是三种相似的EZ-Link NHS-Biotin试剂之一，可在溶液中对抗体、蛋白质和任何其他含有一级胺基的生物分子进行简单有效的生物素化。NHS-Biotin为研究人员提供了优化标记和检测实验的可能性，其中生物素结合的空间阻碍是一个重要因素。由于它是不带电的，并含有简单的烷基空间臂，因此NHS-Biotin化合物是可渗透膜的，并且对于细胞内标记非常有用。

NHS-Biotin是最流行的生物素化试剂类型之一。NHS活化的生物素与碱性缓冲液中的一级氨基（-NH2）高效反应，形成稳定的酰胺键。蛋白质（例如抗体）通常具有几个可用于标记的一级胺基，包括赖氨酸（K）残基的侧链和每个多肽的N-末端。

Reference:
[1]. Fouassier, L., Yun, C. C., Fitz, J. G. and Doctor, R. B. (2000). Evidence for Ezrin-Radixin-Moesin-binding Phosphoprotein 50 (EBP50) Self-association through PDZ-PDZ Interactions. J. Biol. Chem. 275, 25039-25045.
[2]. Nunomura, W., Takakuwa, Y., Parra, M., Conboy, J.G., & Mohandas, N. (2000) Regulation of protein 4.1R, p55 and glycophorin C ternary complex in human erythrocyte membrane. J. Biol. Chem. 275(32): 24540-24546.
[3].Chiu, N.H. & T.K. Christopoulos. 1999. Two-site expression immunoassay using a firefly luciferase-coding DNA label. Clin. Chem. 45: 1954–1959.

Chemical Properties

Cas No.	35013-72-0	SDF
别名	(+)生物素-N-琥珀酰亚胺基酯,Biotin N-hydroxysuccinimide ester, Biotin-OSU, Biotin NHS, Biotin SE
化学名	(2,5-dioxopyrrolidin-1-yl) 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoate
Canonical SMILES	C1CC(=O)N(C1=O)OC(=O)CCCCC2C3C(CS2)NC(=O)N3
分子式	C₁₄H₁₉N₃SO₅	分子量	341.38
溶解度	≥ 17.069mg/mL in DMSO	储存条件	Desiccate at -20°C,protect from light,unstable in solution, ready to use.
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。

溶解性数据

制备储备液
		1 mg	5 mg	10 mg
	1 mM	2.9293 mL	14.6464 mL	29.2929 mL
	5 mM	0.5859 mL	2.9293 mL	5.8586 mL
	10 mM	0.2929 mL	1.4646 mL	2.9293 mL

摩尔浓度计算器
稀释计算器
分子量计算器

计算

动物体内配方计算器 (澄清溶液)

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，注：如该浓度超过该批次药物DMSO溶解度，请先联系GLPBIO）；

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。

注意：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。

Research Update

Availability of NHS-Biotin labeling to identify free protein lysine revealed by experiment and MD simulation

Anal Biochem2018 Sep 15;557:46-58.PMID:30025973DOI:10.1016/j.ab.2018.07.009.

It is known that the crystallizability of protein molecules may be improved by replacing their surface lysine residues with other residue types. Here an experimental method to identify surface lysine residues by NHS-Biotin chemical modification combined with MALDI-TOF MS was proposed and was evaluated using PH1033 protein from Pyrococcus horikoshii. Interestingly, the biotinylation experiment with a protein-reagent molar ratio of 1:1 revealed that only seven of twenty-two lysine residues in the protein comprising 144 residues were labeled. To investigate the result, we analyzed structures from a molecular-dynamics simulation mimicking the experiment. A logistic regression analysis revealed that the biotinylation was significantly correlated with four factors relevant to the local environment of lysine residues: the solvent accessibility, the electrostatic energy, the number of hydrogen bonds, and the estimated pKa value. This result is overall in agreement with that from the same analysis on the crystal structure. However, reflecting the flexibility of the protein molecule in solution state, the factors except for the electrostatic energy were highly variable in the MD structures depending upon the protonation state of Tyr87. The present procedure of biotin-labeling can avoid lysine residues with extensive intramolecular interactions that are incompatible with the rational design of protein crystals.

Biotin as a probe of the surface of Ascaris suum developmental stages

Mol Biochem Parasitol1990 Jun;41(1):45-52.PMID:2385267DOI:10.1016/0166-6851(90)90095-4.

Sulfo-NHS-biotin (aqueous soluble) and NHS-Biotin (organic soluble) labeled similar SDS-2ME (sodium dodecyl sulfate/beta-mercaptoethanol) soluble cuticular proteins of second stage larvae (L2) and third stage Ascaris suum larvae (L3). Comparable analysis of biotin-labeled fourth stage larvae (L4), young adults, and mature adult Ascaris suum revealed strong labeling of several SDS-2ME soluble cuticular proteins with NHS-Biotin, while sulfo-NHS-biotin appeared to strongly label a single SDS-2ME soluble cuticular protein. Both biotin probes labeled only cuticular proteins, since no evidence of internal labeling was observed in any developmental stage examined by either electroblot analysis or by electron microscopy. Our data suggest a greater cuticular permeability to the organic soluble biotin reagent in the later developmental stages (greater than L3) of A. suum than to the aqueous soluble biotin reagent, and may indicate the presence of a hydrophobic barrier in the cuticle of the later stages of the parasite.

Assessment of solvent residues accessibility using three Sulfo-NHS-biotin reagents in parallel: application to footprint changes of a methyltransferase upon binding its substrate

J Mass Spectrom2008 Mar;43(3):360-70.PMID:17968972DOI:10.1002/jms.1328.

NHS-Biotin modification as a specific lysine probe coupled to mass spectrometry detection is increasingly used over the past years for assessing amino acid accessibility of proteins or complexes as an alternative when well-established methods are challenged. We present a strategy based on usage in parallel of three commercially available reagents (Sulfo-NHS-biotin, Sulfo-NHS-LC-biotin, and Sulfo-NHS-LC-LC-biotin) to efficiently assess the solvent accessibility of amino acids using MALDI-TOF mass spectrometry. The same qualitative pattern of reactivity was observed for these three reagents on the THUMPalpha protein at four reagent/polypeptide molar ratios (2 : 1, 6 : 1, 13 : 1, and 26 : 1). Peptide assignment of the detected ions gains in accuracy because of the triple redundancy due to specific increments of monoisotopic mass. These reagents are a good alternative to isotope labeling when using only a single MALDI-TOF mass spectrometer. We observed that hydroxyl groups of serine and tyrosine residues were also modified by these Sulfo-NHS-biotin reagents. The low amount of protein required and the method's simplicity make this procedure accessible and affordable in order to obtain topological information on proteins difficult to purify. This method was used to identify two lysine residues of the TrmG10 methyltransferase from Pyrococcus abyssi that were differentially reactive, modified in the protein but not in the tRNA-protein complex.

NHS-Biotin

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