计算溶液所需的质量、体积或浓度。
ActiBioPure™,EnzymoPure™,His标签,生物活性,高性能 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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。
Endo A 全称糖苷内切酶 A ,基因来源原玻璃蝇节杆菌(Arthrobacter protophormiae),是一种高度特异性糖苷内切酶,可以从糖蛋白上切除没有核心岩藻糖基化的高甘露糖、杂交型 N-聚糖 ,本产品活性高,稳定性好,可以用于抗体糖基化修饰和分析。
.png)
组分和说明
|
使用前准备:
反应试剂:将-20°C储存的10×Glycoprotein Denaturing Buffer缓冲液、10×Glyco Buffer 2及10%NP-40溶液取出,待用。
注意:根据实验需要,准备相应的试剂,如非变性条件下进行去糖基化反应,不需要准备10×Denaturing缓冲液及10%NP-40溶液。
使用方法:
变性条件下糖蛋白去糖基化
1、用去离子水溶解 1-20 µg 的糖蛋白,加入 1 µl 的 10×Glycoprotein Denaturing Buffer,用去离子水定容到 10 µl;
2、75℃孵育 10 min;
3、加入 2 µl 的 10×GlycoBuffer 2,2 µl 的 10% NP-40,轻轻吹打混匀;
4、加入 1-4 µl 的 Endo A,加去离子水到 20 µl,轻轻吹打混匀;
5、37℃ 孵育 1-4hr;
6、用于 SDS-PAGE 分析或 HPLC 分析。
非变性条件下糖蛋白去糖基化
1、取 10-100µg 糖蛋白溶液,加入 2µl 10×GlycoBuffer 2,2-5µl Endo A ,补加纯化水使得反应体系总体积为20µl,轻柔混匀;
2、37℃条件下反应 4-24hr。
注意:当对天然糖蛋白去糖基化时,建议将等量糖蛋白样品进行变性后再同步进行酶切实验作为阳性对照,以确定非变性条件下去糖基化反应的程度。
【注】体系放大时候,孵育时间或酶量需要根据实际情况调整,可低速震荡混匀,可以增加转化率。
注意事项
为了您的安全和健康,请穿实验服并戴一次性手套操作。本产品仅作科研用途!
Endo A, full name Endo-β-N-acetylglucosaminidase A, is derived from Arthrobacter protophormiae. It is a highly specific endoglycosidase that cleaves high-mannose and hybrid-type N-glycans without core fucosylation from glycoproteins. This product features high enzymatic activity and excellent stability, suitable for antibody glycosylation modification and analysis.
.png)
Components List
|
Pre-experiment Preparation
Reaction Reagent:Take out 10× Glycoprotein Denaturing Buffer, 10× GlycoBuffer 2 and 10% NP-40 solution stored at -20°C for standby use.
Note: Prepare corresponding reagents according to experimental requirements. If deglycosylation is performed under non-denaturing conditions, 10× Denaturing Buffer and 10% NP-40 solution are not required.
Operating Protocols
Glycoprotein Deglycosylation under Denaturing Conditions
1. Dissolve 1–20 µg glycoprotein in deionized water, add 1 µL 10× Glycoprotein Denaturing Buffer, and make up to 10 µL with deionized water.
2. Incubate at 75°C for 10 min.
3. Add 2 µL 10× GlycoBuffer 2 and 2 µL 10% NP-40, mix gently by pipetting.
4. Add 1–4 µL Endo A, top up to 20 µL with deionized water and mix gently by pipetting.
5. Incubate at 37°C for 1–4 hr.
6. The reaction mixture can be subjected to SDS-PAGE or HPLC analysis.
Glycoprotein Deglycosylation under Non-denaturing Conditions
1. Take 10–100 µg glycoprotein sample solution, add 2 µL 10× GlycoBuffer 2 and 2–5 µL Endo A. Add purified water to bring the total reaction volume to 20 µL, and mix gently.
2. Incubate the reaction mixture at 37°C for 4–24 hr.
Note: When deglycosylating native glycoproteins, it is recommended to prepare an equal amount of denatured sample for parallel enzymatic digestion as a positive control, to evaluate the deglycosylation efficiency under non-denaturing conditions.Remark: When scaling up the reaction system, adjust incubation time or enzyme dosage as appropriate. Gentle shaking during incubation can improve digestion conversion rate.
Precautions
For personal safety and health, wear a lab coat and disposable gloves during operation. This product is for research use only.
通过匹配包装上的批号来查找并下载产品的 COA,每批产品都进行了严格的验证,您可放心使用!
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | rp223107 | |
| 分析证书 | rp223107 | |
| 分析证书 | rp223107 |
我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。
查看 Bioactive 级别指南 → 查看 ActiBioPure™ 级别指南 → 查看 High Performance 级别指南 → 查看 EnzymoPure™ 级别指南 → 查看 His Tag 级别指南 →