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FITC-BIOTIN 熒光素標(biāo)記生物素的參考文獻(xiàn)

FITC-BIOTIN 是由異硫氰酸熒光素（FITC）和生物素通過(guò)共價(jià)鍵連接而成的熒光探針。該分子保留了生物素對(duì)親和素/鏈霉親和素的高親和力，同時(shí)具有 FITC 的典型熒光特性（激發(fā)波長(zhǎng)約495 nm，發(fā)射約519 nm）。FITC-BIOTIN 常用于免疫熒光、細(xì)胞成像、親和標(biāo)記、ELISA 檢測(cè)及微陣列技術(shù)中。其結(jié)構(gòu)穩(wěn)定，水溶性良好，既具靶向識(shí)別能力又具熒光示蹤功能，是分子探針和納米載體修飾的優(yōu)選標(biāo)記分子。

一、簡(jiǎn)介

名稱：FITC-Biotin

中文名稱：熒光素標(biāo)記生物素

英文別名：Fluorescein-Biotin，F(xiàn)luorescein Isothiocyanate-Biotin，Biotin-FITC

性狀：黃色粉末

結(jié)構(gòu)：

應(yīng)用說(shuō)明：

生物素可用于：少突膠質(zhì)細(xì)胞培養(yǎng)。作為芽孢桿菌(Bacillus)生長(zhǎng)的維生素補(bǔ)充劑。免疫組織學(xué)操作方案中的內(nèi)源性生物素阻斷。生物素是一種小分子，而且它與生物活性大分子共存時(shí)并不會(huì)影響到大分子的生物學(xué)功能。生物素-（鏈霉）親和素系統(tǒng)是多種應(yīng)用所必需的。生物素還可作為各種羧化酶的輔因子/輔酶，如丙酮酸羧化酶、乙酰輔酶A羧化酶 1 和 2、3-甲基巴豆酰輔酶A羧化酶 (MCC) 和丙酰輔酶A羧化酶 (PCC)。它可催化丙酮酸和CO2形成草酰乙酸。與親和素或鏈霉親和素偶聯(lián)的生物素可幫助將靶分子（抗體、核苷酸、蛋白A等）與標(biāo)記系統(tǒng)（酶、熒光、化學(xué)發(fā)光探針）相連接。該復(fù)合物可用于多種檢測(cè)系統(tǒng)，如免疫檢測(cè)、DNA雜交檢測(cè)、免疫組化和流式細(xì)胞術(shù)。該方法還可用于多種目標(biāo)分子的純化和表征。生物素參與基因、細(xì)胞信號(hào)傳導(dǎo)和染色質(zhì)結(jié)構(gòu)的表觀遺傳調(diào)控。熒光素標(biāo)記的生物素(FITC-Biotin)載體酶切:將表達(dá)質(zhì)粒用限制性內(nèi)切酶(同引物的酶切位點(diǎn))進(jìn)行雙酶切,酶切產(chǎn)物行瓊脂糖電泳后,用膠回收Kit或凍融法回收載體大片段。

參考文獻(xiàn)：

來(lái)源：

https://pubs.acs.org/doi/abs/10.1021/nn901474y

主要描述：

We describe molecular capturing properties of protein nanotubes with a controllable ligand binding affinity and size selectivity. These practical biocylinders were prepared using an alternating layer-by-layer (LbL) assembly of protein and oppositely charged poly(amino acid) into the nanoporous polycarbonate (PC) membrane (pore diameter, 400 nm), with subsequent dissolution of the template. The tube wall typically comprises six layers of poly-l-arginine (PLA) and human serum albumin (HSA) [(PLA/HSA)3]. Use of high molecular weight PLA (Mw = ca. 70?000) yielded robust nanotubes, which are available as lyophilized powder. The (PLA/HSA)3 nanotubes swelled considerably in water, although the outer diameter was almost unaltered. Uranyl ion, 3,3′-diethylthiacarbocyanine iodide, and zinc(II) protoporphyrin IX (ZnPP) were bound to the HSA component in the cylinder wall. Similar nanotubes comprising recombinant HSA mutant [rHSA(His)], which has a strong binding affinity for ZnPP, captured this ligand more tightly. Furthermore, addition of excess myristic acid released ZnPP from the tubes through a ligand replacement reaction. The hybrid nanotubes bearing a single avidin layer as an internal surface captured FITC-biotin efficiently. Biotin-labeled nanoparticles are also incorporated into the tubes when their particle size is sufficiently small to enter the pores. Subsequent TEM observation revealed a line of loaded nanoparticles (100 nm) in the one-dimensional space interior.