SMCC Activated Allophycocyanin (SMCC-APC)

SMCC 活化别藻蓝蛋白 (SMCC-APC)

S750587-1mg

1mg

阿拉丁的SMCC Activated Allophycocyanin ，即SMCC活化别藻蓝蛋白，也称SMCC Activated APC，由4-(N-马来酰亚胺基甲基)环己烷-1-羧酸琥珀酰亚胺酯(Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate, SMCC)修饰别藻蓝蛋白形成，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。APC是从螺旋藻(一种蓝绿藻)中分离出来的藻胆蛋白，分子量约为104kDa，具有极高的量子产率和吸收率，可以呈现出强烈且稳定的荧光，易于其它分子交联，非特异性吸附少，灵敏度比常规有机荧光团高100倍以上，因而在生物检测中被广泛用作荧光标签1,2]。APC最大激发波长(Ex)为652nm，荧光最大发射波长(Em)为662nm。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的条件下NHS酯基团与APC的伯氨基反应形成酰胺键，而马来酰亚胺基团在pH6.5-7.5的条件下可以与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APC与其它含巯基的分子如蛋白等的偶联3]，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 Allophycocyanin 与蛋白质的偶联原理图。本产品为APC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-APC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 Allophycocyanin is formed by modifying Allophycocyanin (APC) 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.APC is a phycobiliprotein isolated from spirulina (a type of blue-green algae). It has a molecular weight of approximately 104kDa and exhibits a very high quantum yield and absorbance, resulting in strong and stable fluorescence. It is easily cross-linked with other molecules and has minimal non-specific adsorption. Its sensitivity is more than 100 times higher than conventional organic fluorophores, making it widely used as a fluorescent label in biological assays [1,2]. The maximum excitation wavelength (Ex) of APC is 652nm, and the maximum emission wavelength (Em) is 662nm.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 APC 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 APC with other molecules containing thiol groups [3] (Figure 1). The cyclohexane in SMCC structure greatly reduces the hydrolysis rate of the maleimide group, allowing SMCC-modified proteins to be stored in lyophilized form.Figure 1. Schematic diagram of the conjugation of Aladdin's SMCC Activated Allophycocyanin to proteins.This product is a purified conjugate of Allophycocyanin (APC) 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.This 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.References 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 Allophycocyanin should also be stored at 4℃ in the dark. Do not freeze. ØAvoid 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. Instructions for Use ： none