本产品是唾液酸酶（α2- 3,6,8,9）和β1- 4半乳糖苷酶两种E.coli表达的重组酶的混合物。其中，唾液酸酶（α2- 3,6,8,9）来源于Arthrobacterureafaciens，可以水解糖蛋白和寡聚糖上通过α2- 3，α2- 6，α2- 8和α2- 9键连接的唾液酸，其水解α2- 3和α2- 6键的速度要大于α2- 8和α2- 9键。β1- 4半乳糖苷酶来源于Streptococcus pneumoniae，是一种具有高度特异性的糖苷外切酶，催化水解糖蛋白和寡糖末端的非还原的半乳糖残基。通过两种酶的协同作用，本产品可以去除糖蛋白的唾液酸和非还原的半乳糖残基，使得糖蛋白糖型向G0/G0F转变。
酶活定义
唾液酸酶（α2- 3,6,8,9） ：
一个单位定义为在 37°C ，5分钟内，在10ul的总反应体积中，从1nmol Neu5Acα2- 3Galβ1- 3GlcNAcβ1- 3Galβ1- 4Glc- 7- amino- 4- methyl- coumarin（AMC）中切割解 >95% 的末端α- Neu5Ac所需的酶量。
β1- 4半乳糖苷酶：
一个单位是指10ul反应体系中， 37°C 条件下，1小时内将1nmol Galβ1- 4GlcNAcβ1- 3Galβ1- 4Glc- 7- amino- 4- methyl- coumarin中 >95% 的末端 β‑D‑半乳糖 水解所需要的酶量。
本品含量
| 酶 | 5rxns | 20rxns
|
| 唾液酸酶（α2- 3,6,8,9） | 100U/支 | 400U/支 |
β1-4半乳糖苷酶
| 250U/支 | 1000U/支 |
组分表
| 储运单元货号 | 储运单元名称 | 组分货号 | 组分名称 | 物理外观 | 5rxns | 20rxns | 储存 |
| P1527895WT | 蛋白脱糖混合液I(2-8℃组分) | P1527895A | 蛋白脱糖混合液I | 液体 | 25µl | 100µl | 2-8℃. |
| P1527895DT | 蛋白脱糖混合液I (-20℃组分) | P1527895B | 10×糖苷酶切反应液 | 液体 | 200µl | 200µl | -20℃ |
注：发货时各组分将按照对应储存温度（2-8°C/-20°C）独立包装，采用差异化温控运输，保障产品最佳性能。若同一订单包含多种温区组分，包裹可能分批次送达，请以实际收货情况为准。收货后，请立即依据组分表标注的温度条件分类存放。所有组分在未拆封、运输及储存条件均符合要求的情况下，有效期为自生产之日起 12 个月
产品应用
去除糖蛋白末端的唾液酸和非还原的半乳糖残基。
使用方法（仅供参考）
①建议按照以下体系进行反应
| 组分 | 体积 |
|---|
| 底物 | 100μg 糖蛋白（10-40μl） |
| 10×糖苷酶切反应液 | 5.0μl |
| 蛋白脱糖混合液 I | 5.0μl |
| 水 | 补足总体积 50μl |
②25°C 保温30分钟，再 37°C 保温16小时；
③65°C ，20分钟可以热失活蛋白脱糖混合液 I 。
注意事项
-本品收到后尽量避免冻融
- 使用时请穿实验服并佩戴一次性手套
- 本品不得直接用于临床诊断和治疗。
This product is a mixture of two recombinant enzymes expressed in E. coli: sialidase (α2‑3,6,8,9) and β1‑4‑galactosidase. Sialidase (α2‑3,6,8,9) is derived from Arthrobacter ureafaciens. It hydrolyzes sialic acids linked via α2‑3, α2‑6, α2‑8 and α2‑9 linkages on glycoproteins and oligosaccharides, with higher hydrolysis rates toward α2‑3 and α2‑6 linkages than α2‑8 and α2‑9 linkages. β1‑4‑galactosidase originates from Streptococcus pneumoniae, a highly‑specific exoglycosidase that catalyzes the hydrolysis of non‑reducing terminal galactose residues on glycoproteins and oligosaccharides. Through the synergistic effect of the two enzymes, this product removes sialic acids and non‑reducing terminal galactose residues from glycoproteins, shifting glycoprotein glycoforms toward G0/G0F.
Enzyme Activity Definition
Sialidase (α2‑3,6,8,9): One unit is defined as the amount of enzyme required to cleave >95 % of terminal α‑Neu5Ac from 1 nmol Neu5Acα2‑3Galβ1‑3GlcNAcβ1‑3Galβ1‑4Glc‑7‑amino‑4‑methyl‑coumarin (AMC) in a total reaction volume of 10 μL at 37 °C within 5 minutes.
β1‑4‑Galactosidase: One unit refers to the amount of enzyme required to hydrolyze >95 % of terminal β‑D‑galactose from 1 nmol Galβ1‑4GlcNAcβ1‑3Galβ1‑4Glc‑7‑amino‑4‑methyl‑coumarin in a 10 μL reaction system at 37 °C within 1 hour.
Content per Vial
| Enzyme | 5 rxns | 20 rxns |
|---|
Sialidase (α2‑3,6,8,9)
| 100U/vial | 400U/vial |
β1‑4‑Galactosidase
| 250U/vial | 1000U/vial |
Component
| Storage Unit No. | Storage Unit Name | Component No. | Component Name | Appearance | 5rxns | 20rxns | Storage |
| P1527895WT | Protein Deglycosylation Mix I(2-8℃ Components) | P1527895A | Protein Deglycosylation Mix | Liquid | 25µl | 100µl | 2-8℃. |
| P1527895DT | Protein Deglycosylation Mix I(-20℃ Components) | P1527895B | 10× Glycosidase Cleavage Reaction Buffer | Liquid | 200µl | 200µl | -20℃ |
Note: During shipment, each component is individually packaged and transported under temperaturecontrolled conditions corresponding to its storage requirement ( 2-8°C/-20°C ) to maintain optimal product performance. For a single order containing components requiring different storage temperatures, packages may be delivered in separate shipments, subject to actual receipt. Upon receipt, store all components immediately according to the temperature specifications listed in the component list. The shelflife of all unopened components is 12 months from the date of manufacture, provided that both shipping and storage conditions are strictly followed.
Product Applications
Removal of terminal sialic acids and non‑reducing terminal galactose residues from glycoproteins.
Procedure (For Reference Only)
①It is recommended to set up the reaction system as follows:
| Component | Volume |
|---|
| Substrate | 100 μg glycoprotein (10‑40 μL) |
| 10× Glycosidase Reaction Buffer | 5.0 μL |
| Protein Deglycosylation Mix I | 5.0 μL |
| Water | Adjust to final volume 50 μL |
②Incubate at 25 °C for 30 min, followed by incubation at 37 °C for 16 h.
③Heat‑inactivate Protein Deglycosylation Mix I at 65 °C for 20 min.
Notes
‑ Avoid repeated freeze‑thaw cycles upon receipt of this product.
‑ Wear laboratory coat and disposable gloves during operation.
‑ This product is not intended for clinical diagnosis or therapeutic use.
危险品化学品经营许可证（带存储）