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阿拉丁生产的LumiDye™ SMCC Activated HRP (SMCC-HRP)，即SMCC活化辣根过氧化物酶，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)修饰辣根过氧化物酶(Horseradish Peroxidase, HRP)形成，每分子HRP通常含多个马来酰亚胺基团，同时保留大部分HRP活性，可直接偶联任何含巯基化合物，无需进一步修饰。马来酰亚胺与巯基的相互作用高度特异且高效，形成共价键，可作为HRP探针用于ELISA、Western Blot等检测体系。
本产品为HRP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-HRP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-HRP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-HRP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10mg/mL，最佳标记质量比为IgG:HRP=1:1，即摩尔标记比为IgG:HRP=1:3.5，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。
图1 LumiDye™ SMCC Activated HRP (SMCC-HRP)（M1491737）与蛋白质的偶联原理图
保存条件
-20ºC避光保存，12个月内有效。
注意事项
1. 收到本产品后请立即使用或按照说明书推荐的条件保存。随着保存时间的延长，Activated HRP的酶活力和标记能力可能会有一 定程度的下降；
2. 待标记的抗体或蛋白最好溶解于无氨的缓冲液中（如：MES，MOPS，HEPES，PBS），pH 为 6.5-8.5，且溶液中无 NaN3 等防腐剂（若含有此类物质需先纯化后，再标记）；
3. 操作过程中，务必穿戴实验服、一次性手套及防护装备；
4. 所有产品仅限于研究用途。
LumiDye™ SMCC Activated HRP (SMCC-HRP) is formed by modifying Horseradish Peroxidase (HRP) with Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC). SMCC-HRP generally contains several maleimide groups per HRP molecule while maintaining most of the HRP activity. After conjugation, the activated HRP will form a covalent interaction with the peptides or ligands containing sulfhydryl groups and can be used as a HRP probe for detection in different assays such as ELISA and western blot etc.
This product is a purified conjugate of HRP 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 10 mg/mL. For IgG labeling, the recommended mass ratio is IgG:HRP = 1:1, which corresponds to a molar labeling ratio of IgG:HRP = 1:3.5. Other proteins may be labeled using different molar ratios as needed.Note: The SMCC-HRP solution must be prepared fresh before each use.
Figure 1. Schematic diagram of the conjugation of LumiDye™ SMCC Activated HRP (SMCC-HRP) (M1491737）to proteins.
Storage
Store at -20℃ in the dark for up to 12 months.
Matters needing attention
1. Upon receipt of this product, it should be used immediately or stored according to the recommended conditions. With prolonged storage, the enzymatic activity and labeling capability of the Activated HRP may decrease to some extent.
2. The antibody or protein to be labeled should be dissolved in an amine-free buffer (e.g., MES, MOPS, HEPES, PBS) at a pH of 6.5–8.5. The solution must be free of preservatives such as sodium azide. If present, these must be removed via purification prior to the labeling reaction.
3. During operation, always wear a lab coat, disposable gloves, and protective equipment.
4. All products are for research use only.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0332695 |
分析证书 | M1491737 |
| ZJ26F0332696 |
分析证书 | M1491737 |
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