Biotin-dPEG?4-hydrazide, product number QBD-10219, is a hydrophilic biotinylation reagent designed for labeling the carbohydrate coats of glycoproteins with biotin. The amphiphilic spacer provides excellent water solubility for poorly water-soluble biotin. Moreover, the dPEG?4 spacer is longer than the hydrophobic aminocaproic acid (LC) spacer used in biotin-LC-hydrazide, so it provides somewhat better access to the biotin binding pockets of avidin and streptavidin. Additionally, the dPEG?4 spacer reduces or eliminates non-specific binding in many applications.nBiotin-dPEG?4-hydrazide, QBD-10219, is a carbonyl-reactive compound that forms semi-permanent hydrazone bonds with aldehydes, ketones, and carboxylic acid groups. Although there are numerous natural small molecules containing aldehydes and ketones, macromolecules of biological importance (peptides, proteins, and nucleic acids) rarely contain them. However, glycosylated proteins such as antibodies frequently contain carbohydrate residues known as reducing sugars that can be oxidized with sodium periodate, neuraminidase, or galactose oxidase to form aldehydes.nThe aldehydes thus formed then can be reacted with Biotin-dPEG?4-hydrazide at pH 5 – 7, labeling the protein with biotin through the formation of a Schiff base. Aniline or para-phenylenediamine catalyzes the reaction, forming the Schiff base quickly. For additional bond stability, sodium cyanoborohydride can be used to reduce the Schiff base to a secondary amine. Via carbodiimide chemistry (for example, 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, also known as EDC), Biotin-dPEG?4-hydrazide reacts with carboxylic acid groups forming a stable bond.nThe amphiphilic dPEG?4 spacer in Biotin-dPEG?4-hydrazide provides the ability to use Biotin-dPEG?4-hydrazide directly in water or aqueous buffer, making it more biocompatible than LC-biotin. Moreover, QBD-10219 will not trigger protein or peptide aggregation or cause non-specific binding due to hydrophobicity issues from the linker, both of which can occur with traditional biotin linkers like LC-biotin.nBiotin-dPEG?4-hydrazide is highly useful for installing a biotin label onto glycoproteins. It is also used in cell labeling to label glycoproteins on cell surfaces. Biotinylated glycoproteins can be identified using streptavidin-horseradish peroxidase (HRP) conjugates, because of the high affinity between biotin and streptavidin. Biotinylated secondary antibodies are used frequently with avidin or streptavidin in plate-based assays such as ELISA and western blots. This reagent, coupled with avidin that has been modified to have reduced biotin-binding affinity, is for affinity purification of molecules.
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Click Chemistry Tools(CCT)公司位于美國亞利桑那州斯科茨代爾(Scottsdale,AZ)。自公司成立以來,一致致力于不斷開發(fā)和擴展一系列連接產(chǎn)品。他們利用特別新的研究成果研發(fā)生產(chǎn)出新穎的交聯(lián)和標記試劑。產(chǎn)品被廣泛用于藥物研發(fā)、癌癥治療以及各類診斷技術(shù)等。此外,CCT還可以提供個性化的定制產(chǎn)品,已滿足不同用戶的需求。目前CCT的產(chǎn)品線主要包括:Azides(Biotin-Azides)試劑、Bis-sulfone交聯(lián)試劑、DBCO試劑、TCO試劑、Terminal Alkyne試劑、Tetrazine試劑、標記試劑盒以及分選用底物等。nFluoroprobes公司位于美國亞利桑那州斯科茨代爾(Scottsdale, AZ),自成立以來,一直致力于不斷開發(fā)一系列的連接(linker)產(chǎn)品。利用特別新的研究成果研發(fā)生產(chǎn)出各類新穎的交聯(lián)和標記試劑。其產(chǎn)品被廣泛用于藥物研發(fā)與篩選、疾病治療方法和診斷技術(shù)等領(lǐng)域。此外,F(xiàn)luoroprobes還可以根據(jù)客戶的個性化需求提供各類定制連接產(chǎn)品。代表產(chǎn)品有Classic Fluorescent Dyes, AZDye, Cyanine Dyes, IR Dyes, MB dyes以及一些標記試劑盒等。nQuanta Biodesign公司位于美國中部的俄亥俄州,成立于1997年。也是專注于生產(chǎn)研發(fā)各類標記產(chǎn)品的品牌之一。其研發(fā)的產(chǎn)品與dPEG技術(shù)相結(jié)合,生產(chǎn)出他們獨有的專利產(chǎn)品dPEG Building Blocks and Linkers,被廣泛用于細胞治療和分子診斷等研究領(lǐng)域。另外,還可為廣大科研工作者提供多種化學修飾試劑以及傳統(tǒng)Liner等。n目前上述三個品牌都已被被Vectorlabs收購并整合到Vectorlabs產(chǎn)品中