SMCC Activated R-Phycoerythrin (SMCC-RPE)

SMCC 活化 R-藻红蛋白 (SMCC-RPE)

S750588-1mg

1mg

阿拉丁生产的SMCC Activated R-Phycoerythrin ，即SMCC活化R-藻红蛋白，也称SMCC Activated RPE、SMCC Activated R-PE、SMCC-PE、SMCC活化藻红蛋白，是由4-(N-马来酰亚胺基甲基)环已烷-1-羟酸琥珀酰亚胺酯(Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate, SMCC)修饰R-藻红蛋白(R-Phycoerythrin, R-PE/RPE)形成，可与含巯基的分子如蛋白等偶联，偶联物常用于荧光显微检测、荧光免疫测定、双色或多色荧光分析、癌细胞表面抗原检测、流式细胞荧光测定等抗体荧光标记以及活体成像等诊断及生物工程技术。RPE是一种从红藻中分离出来的具有强烈荧光的藻胆蛋白，分子量约为240kDa。RPE具有极高的量子产率，呈现出极亮的红橙色荧光，不易淬灭，保存期长，水溶性极佳，与常规有机荧光团相比更明亮、更稳定1]，因而在生物检测中被广泛用作荧光标签。RPE最大激发波长(Ex)为565nm，次级激发波长为480nm和545nm，荧光最大发射波长(Em)为575nm。SMCC是一种常见的胺-巯基交联剂，含一个胺反应性N-羟基琥珀酰亚胺(NHS酯)基团和巯基反应性马来酰亚胺基团，在pH7-9条件下NHS酯基团与RPE的伯氨基反应形成稳定的酰胺键，而马来酰亚胺基团可以与在pH6.5-7.5条件下另一分子如蛋白等的巯基反应形成稳定的硫醚键，从而实现RPE与其它含巯基的分子如蛋白等的偶联2]，其原理如图1所示。因SMCC结构中含有环己烷，环己烷大大降低了马来酰亚胺基团的水解速率，使SMCC修饰过的蛋白质可以通过冻干的形式储存。图1.阿拉丁SMCC Activated R-Phycoerythrin 与蛋白质的偶联原理图。本产品为RPE与SMCC的连接产物，经纯化并去除过量的SMCC，溶于PBS，最终以冻干粉的形式提供，长期储存也不损失巯基反应性，保证了SMCC-RPE与其它含巯基分子如蛋白质等偶联的最佳性质。本产品可以针对抗体等生物大分子进行偶联，这些分子具有半胱氨酸，而半胱氨酸上有巯基。当半胱氨酸上的巯基形成二硫键时，可以通过还原剂处理以产生游离的巯基基团。当蛋白等生物大分子具有可用的伯胺基团时，也可以通过化学反应添加所需的巯基基团。4ºC避光保存，一年有效。切勿 Aladdin's SMCC Activated R-Phycoerythrin is formed by modifying R-Phycoerythrin (R-PE/RPE) with Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate . It contains a maleimide group that reacts with thiol groups of target molecules, such as proteins. This product is commonly used in fluorescence microscopy, fluorescence immunoassays, dual or multicolor fluorescence analysis, cancer cell surface antigen detection, flow cytometry fluorescence assays, antibody fluorescence labeling, as well as diagnostics and biotechnology applications such as live cell imaging.R-Phycoerythrin (RPE) is a highly fluorescent allophycocyanin protein isolated from red algae, with a molecular weight of approximately 240kDa. RPE has a high quantum yield and exhibits intense red-orange fluorescence that is resistant to quenching and has a long shelf life. It is highly water-soluble and more stable than conventional organic fluorophores [1], making it widely used as a fluorescent label in biological assays. RPE's primary absorption peak is at 565nm with secondary peaks at 480nm and 545nm. Its maximum emission wavelength (Em) is 575nm.SMCC is a common amine-thiol cross-linking reagent that contains a reactive N-hydroxysuccinimide (NHS ester) group and a maleimide group reactive with thiol groups. Under pH 7-9, the NHS ester group reacts with the primary amino groups of RPE to form stable amide bonds, while the maleimide group can react with thiol groups of other molecules, such as proteins, under pH 6.5-7.5 to form stable thioether bonds, enabling the conjugation of RPE to other molecules containing thiol groups [2] (Figure 1). The cyclohexane in SMCC structure greatly reduces the hydrolysis rate of maleimide group, allowing SMCC-modified proteins to be stored in lyophilized form.Figure 1. Schematic diagram of the conjugation of Aladdin's SMCC Activated R-Phycoerythrin to proteins.This product is a purified conjugate of R-Phycoerythrin (RPE) and SMCC. It is provided as a lyophilized powder, and can be dissolved in PBS. The long-term storage of this product does not affect its thiol reactivity, ensuring the optimal properties for conjugation with other thiol-containing molecules such as proteins. ØThis product is chemically activated for easy conjugation to biomacromolecules such as antibodies. These molecules contain cysteine residues. Disulfide bonds formed between cystein residues can be treated with a reducing agent, such as DTT and β-mercaptoethanol, to release free thiol groups. Thiol groups can also be added to primary amine groups of biomacromolecule, such as a protein, through chemical reactions. Precautions ： Fluorescent dyes are prone to quenching. Please store and use them in the dark to slow down fluorescence quenching.Products conjugated with the SMCC Activated R-Phycoerythrin should also be stored at 4℃ in the dark. Do not freeze.Avoid using buffers containing primary amines and thiols during the reaction to minimize unnecessary competitive reactions.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.References:1. Batard P, Szollosi J, Luescher I, Cerottini JC, MacDonald R, et al. 2002. 48(2):97-105.2. Li Y, He Q, Hu X, Liu Y, Cheng X, et al. Biomater Sci Polym Ed. 2017. 28(7):629-647. Instructions for Use ： none