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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
阿拉丁生产的LumiDye™ SMCC Activated Streptavidin (SMCC-SA)，即SMCC活化链霉亲和素，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)修饰链霉亲和素(Streptavidin，SA)形成，可与含巯基的分子如蛋白等偶联，偶联物无需额外纯化，可直接用于蛋白质印迹、ELISA及免疫组化等应用。
链霉亲和素是一个非糖基化的蛋白质。它对生物素具有优异的高亲和力，并且在大幅度的pH和温度范围内稳定。由于其对生物素的高亲和力，它被用来桥接生物素化的分子。每个链霉亲和素分子可以结合4个生物素分子。链霉亲和素是一种来源于阿维丁链霉菌（streptomyces avidinii）的生物素结合蛋白质，其非特异性结合远比亲和素低，链霉亲和素蛋白具有在中性pH值几乎没有净电荷，不含有碳水化合物等更有利的化学性质，所以它被广泛用来作为抗生物素蛋白的替代品。
本产品为SA与SMCC的连接产物，经纯化并去除过量的SMCC，溶于含3%海藻糖的PBS-EDTA中，SMCC-SA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-SA与其它含巯基分子如蛋白质等偶联的最佳性质。
使用方法
SMCC-SA具有巯基反应性，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，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。
本产品规格为0.1 mg和1 mg，最佳使用浓度为5mg/mL(纯水复溶)，最佳标记质量比为IgG:SA=1:1，即摩尔标记比为IgG:SA=1:3，其他蛋白可采用其他摩尔比进行标记。注：本产品需现用现配。
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图1 LumiDye™ SMCC Activated Streptavidin (SMCC-SA)（M1491735）与蛋白质的偶联原理图
保存条件
4ºC避光保存，12个月内有效，切勿-20ºC或更低温度保存。
注意事项
1. 避免在反应过程中使用含有伯胺和巯基的缓冲液，以减少不必要的竞争反应；
2. 本产品的偶联产物，须避光保存，请勿冷冻；
3. 在正式实验前，需将试剂恢复至室温，较低的温度会影响偶联效果。开启瓶盖之前，先适当离心，将粉末聚集于管底，避免开盖时损失。
4. 操作过程中，务必穿戴实验服、一次性手套及防护装备；
5. 所有产品仅限于研究用途。
LumiDye™ SMCC Activated Streptavidin (SMCC-SA) is formed by Streptavidin with Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC). It contains a maleimide group that reacts with thiol groups of target molecules, such as proteins. The conjugate can be used immediately in applications such as Western blot, ELISA, and Immunohistochemistry (IHC) without further purification.
Streptavidin is a non-glycosylated protein. It exhibits exceptionally high affinity for biotin and remains stable across a wide range of pH levels and temperatures. Due to its high affinity for biotin, it is used to bridge biotinylated molecules. Each streptavidin molecule can bind four biotin molecules. Streptavidin is a biotin-binding protein derived from Streptomyces avidinii. Its non-specific binding is significantly lower than that of avidin, and it possesses more favorable chemical properties, such as having almost no net charge at neutral pH and being carbohydrate-free. Therefore, it is widely used as a substitute for avidin.
This product is a purified conjugate of SA and SMCC. After purification to remove excess SMCC, it is dissolved in PBS-EDTA with 3% trehalose and ultimately supplied as a lyophilized powder. 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.
Instruction for use
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.
Two options are available: 0.1 mg and 1.0 mg. The optimal concentration is 5 mg/mL(Reconstitute in ultrapure water). For IgG labeling, the recommended mass ratio is IgG:SA = 1:1, which corresponds to a molar labeling ratio of IgG:SA = 1:3. Other proteins may be labeled using diff erent molar ratios as needed. Note: The SMCC-SA solution must be prepared fresh before each use.
Figure 1. Schematic diagram of the conjugation of LumiDye™ SMCC Activated Streptavidin (SMCC-SA) (M1491735）to proteins.
Storage
Store at 4℃ in the dark for up to 12 months. Do not store at -20℃ or lower temperatures.
Matters needing attention
1. Avoid using buffers containing primary amines and thiols during the reaction to minimize unnecessary competitive reactions.
2 Products conjugated with the SMCC-SA should be stored at 4℃ in the dark. Do not freeze.
3.Before formal experimentation, allow all reagents to reach room temperature, as lower temperatures may impair conjugation efficiency. Before opening the bottle cap, centrifuge it briefly to settle the powder at the bottom of the tube, preventing loss during opening.
4. During operation, always wear a lab coat, disposable gloves, and protective equipment.
5. All products are for research use only.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0333426 |
分析证书 | M1491735 |
| ZJ26F0333427 |
分析证书 | M1491735 |
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