Biotin Hydrazide
(Synonyms: 生物素酰肼) 目录号 : GC12253
A biotinylation reagent
Cas No.:66640-86-6
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Biotinylation method [1]: | |
|
Sample |
human umbilical cord HA |
|
Preparation method |
Soluble in DMSO. Dissolve glycoprotein at a concentration of 1-5 mg/mL in cold 100mM Sodium Acetate, pH 5.5. |
|
Reaction Conditions |
1 mM, overnight at 4℃ |
|
Applications |
One hundred milligrams of human umbilical cord HA was dissolved in 0.1 M Mes, pH 5.0, to a final concentration of 1 mg/ml and allowed to dissolve for at least 24 h at 4℃ prior to the coupling of biotin. Biotin hydrazide was dissolved in DMSO as a stock solution of 100 mM and added to the HA solution to a final concentration of 1 mM. A stock solution of 1-ethyl-3-(3-dimethylamin-opropyl) carbidodiimide (EDAC) was prepared as a 100 mM stock solution in dH2O and added to the HA–biotin solution at a final concentration of 30 mM. This solution was left stirred overnight at 4℃. Unlinked biotin and EDAC were removed after the addition of 4M guanidine–HCl. The dialyzed, biotinylated HA (bHA) was aliquoted and stored at 0207C for up to several months. |
|
References: [1]. Gregory I. Frost and Robert Stern. A Microtiter-Based Assay for Hyaluronidase Activity Not Requiring Specialized Reagents. ANALYTICAL BIOCHEMISTRY 251, 263–269 (1997). |
|
Biotin hydrazide is a biotinyl derivative that can be used as a probe for the determination of protein carbonylation, which is a component of several diseases. Protein carbonylation, an irreversible post translational modification (PTM), is caused by attack of reactive oxygen species (ROS), numerous lipid oxidation products (such as α,β-unsaturated γ-hydroxyalkenals), or nonemzymatic glycation resulting in the loss of protein function. Biotin hydrazide is a preferred carbonyl-reactive probe for its direct reaction and chemistry and no requirement of additional catalysts or reducing agents. Biotin hydrazide exclusively and readily derivatizes carbonyl groups at pH 5.5, which facilitates its use to measure carbonylated proteins in biological samples.
Reference
[1].Kenneth Hensley, Kelly S. Williamson. Protein Carbonyl Determination Using Biotin Hydrazide. Methods in Biological Oxidative Stress. Methods in Pharmacology and Toxicology 2003, pp 195-199
[2].Madian AG, Regnier FE. Proteomic identification of carbonylated proteins and their oxidation sites. J Proteome Res. 2010;9(8):3766-3780.
| Cas No. | 66640-86-6 | SDF | |
| 别名 | 生物素酰肼 | ||
| 化学名 | 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanehydrazide | ||
| Canonical SMILES | C1C2C(C(S1)CCCCC(=O)NN)NC(=O)N2 | ||
| 分子式 | C10H18N4O2S | 分子量 | 258.3 |
| 溶解度 | ≥ 12.9mg/mL in DMSO with gentle warming, ≥ 2.98 mg/mL in Water with ultrasonic | 储存条件 | Store at -20°C,protect from light |
| 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 | 3.8715 mL | 19.3573 mL | 38.7147 mL |
| 5 mM | 0.7743 mL | 3.8715 mL | 7.7429 mL |
| 10 mM | 0.3871 mL | 1.9357 mL | 3.8715 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。
A cleavable biotin tagging reagent that enables the enrichment and identification of carbonylation sites in proteins
Anal Bioanal Chem 2016 Jan;408(3):865-74.26613796 10.1007/s00216-015-9176-2
The utility of a new, cleavable tag for identifying and enriching protein carbonyls is examined. Using a model system, human serum albumin modified with acrolein, the EZ-Link alkoxyamine-PEG4-SS-PEG4-biotin affinity tag, was tested for its ability to label protein carbonyls in proteomic analyses of protein carbonylation. The efficiency of the labeling was assayed and compared to standard Biotin Hydrazide reagents. The label was also tested in liquid chromatography-tandem mass spectrometry (LC/MS/MS) experiments. The quality of the fragmentation spectra was assessed and the relative detection efficiency of various modification sites was compared to standard Biotin Hydrazide reagents. Finally, the viability of using the label with streptavidin bead enrichment protocols in a standard proteomics workflow was probed.
Biotin-conjugated reagents as site-specific probes of membrane protein structure: application to the study of the human erythrocyte hexose transporter
Anal Biochem 1990 Nov 1;190(2):297-303.2127160 10.1016/0003-2697(90)90197-h
A novel labeling procedure using biotin-conjugated protein-modifying reagents has been employed to study the structure and function of the human erythrocyte hexose transporter. The carbohydrate moiety of the isolated, reconstituted transporter was labeled by using galactose oxidase/Biotin Hydrazide. Cysteine residues, which are essential for transporter function, were tagged with a biotin-conjugated maleimide. Labeling with this reagent inhibited the binding of cytochalasin B to the transporter. Following sodium dodecyl sulfate-gel electrophoresis, labeling of the transporter and its proteolytic fragments was detected by Western blotting and probing with alkaline phosphatase-conjugated avidin. After tryptic cleavage of the transporter into two membrane domains, preparations reacted with galactose oxidase/Biotin Hydrazide were labeled on the 25-kDa glycosylated fragment, but not on the carbohydrate-free 19-kDa peptide. Biotin-maleimide-labeled cysteine residues on both peptides. Transporter polypeptide was fragmented more extensively using Staphylococcus aureus V8 protease. Limited digestion produced a broad band of 30-50 kDa and sharper bands of 23 and 21 kDa. More extensive digestion resulted in the disappearance of the 23-kDa peptide and the appearance of sharp bands of 20, 19, 17, 13, 11, 8, and 7 kDa. Biotin label introduced with galactose oxidase/Biotin Hydrazide was found on the broad 30-kDa band, confirming its identity as a glycopeptide. All of the peptides weighing more than 11 kDa contained cysteine residues labeled with biotin maleimide, while the 8- and 7-kDa peptides were unlabeled. These results demonstrate the potential usefulness of biotin-conjugated reagents as site-specific probes of membrane protein structure.
Nonradioactive hybridization probes prepared by the reaction of Biotin Hydrazide with DNA
Biochem Biophys Res Commun 1987 Jan 30;142(2):519-26.3101692 10.1016/0006-291x(87)90305-6
A novel one-step chemical method has been developed for the introduction of biotin into nucleic acids for non-isotopic hybridization. The method is based on the interaction of Biotin Hydrazide with unpaired cytosine residues. The interaction is catalyzed by sodium bisulfite with an optimum at a buffered pH of about 4.5. The reaction reached its maximum after 24 h incubation at a Biotin Hydrazide concentration of 10 mg/ml. Using streptavidin-alkaline phosphatase conjugates, the limits for detecting the biotinylated probe, either adsorbed directly to nitrocellulose or hybridized to filter-bound target DNA, were 0.3 and 0.9 pg, respectively. The salience of the approach described here over previously used biotin derivatives is that it is quick (one-step), simple and does not involve any enzymatic or instrument-mediated step to introduce the reporter moiety. In addition, other low- and high-molecular-weight hydrazides (e.g. fluorescent or enzyme hydrazides) can serve as the reporter group. The same procedure may be employed for the single-step biotinylation of free cytidine.
Highly sensitive biotin-labelled hybridization probe
Chem Pharm Bull (Tokyo) 1989 Jul;37(7):1831-3.2805162 10.1248/cpb.37.1831
A simple chemical method for introducing biotin into nucleic acids has been developed for the synthesis of nonisotopic hybridization probes. The method is based on the reaction of Biotin Hydrazide with amino residues of nucleic acids by using glutaraldehyde as a bifunctional coupling reagent. Biotin-labelled deoxyribonucleic acid (DNA) was detected by the use of alkaline phosphatase-labelled avidin, and alkaline phosphatase activity was measured by colorimetric and chemiluminescence methods. The chemiluminescence method using the nicotinamide adenine dinucleotide phosphate (NADP)/alcohol/alcohol dehydrogenase/microperoxidase/isoluminol system gave the highest sensitivity. A few picograms of lambda-phage DNA coated on a microtiter plate well could be detected by this method.
Comparative electrochemical behaviour of Biotin Hydrazide and photobiotin. Importance in the development of biosensors
Biosens Bioelectron 1999 Dec;14(8-9):729-35.10641292 10.1016/s0956-5663(99)00049-4
The cyclic voltammetric behaviour of Biotin Hydrazide and photobiotin on carbon paste electrodes has been studied. Biotin Hydrazide presents an anodic and irreversible process, meanwhile photobiotin presents two, adsorptive in nature. This characteristic makes photobiotin desirable for following the interaction between biotin and streptavidin, being possible to detect a streptavidin concentration of 10(-12) M. The evidence of this reaction has been shown either directly in solution or on the electrode surface. Photobiotin as the molecule portable of analytical information and carbon paste as the solid support could be applied to the development of sensors based on the oxidation of this molecule.