计算溶液所需的质量、体积或浓度。
酶增效表面活性剂
有货
为什么选择此级别
BioReagent 级 提供 ≥95%(NMR) 纯度，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
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。
文献证明
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。
概述
酶增效表面活性剂是一种阴离子型温和蛋白质变性剂，专为提升溶液内蛋白酶（如胰蛋白酶、Lys-C、PNGase F、Asp-N等）的水解效率而设计。它通过促进蛋白质的溶解与结构展开，使内切蛋白酶更易接近疏水性蛋白质的酶切位点，从而显著提高裂解效率并大幅缩短酶解时间。
与常规变性剂不同，该表面活性剂在终浓度低于1%时几乎不影响上述水解酶的活性，同时可避免对目标蛋白引入氧化、脱酰胺等非期望修饰，且与质谱检测完全兼容。
得益于其温和而高效的变性特性，它还能改善疏水性蛋白质及肽段的溶解度，减少昂贵蛋白酶的用量，助力实验室在更短时间内完成高回收率的酶解流程，满足高通量分析的需求。
产品应用
提高蛋白水解酶的裂解效率，缩短酶切时间
使用方法（仅供参考）
使用前将本品10000g离心10s（仅针对1mg和3mg规格），确保所有粉末都在管底部。
1.PNGase F酶切-非还原条件
① 用10×糖苷酶切反应液（500mM磷酸二氢钠，pH=7.5）将酶增效表面活性剂 复溶至终浓度10mg/ml，轻柔混匀。
②用去离子水溶解50-100μg糖蛋白，加入5μl含本品的10×糖苷酶切反应液，并用去离子水定溶至48-49μl。
③80-90°C加热3-5min，冷却至室温（此步骤可选，但加热的酶切效果更佳）。
④加入1-2μl的PNGase F，使得反应总体积为50μl，混匀。
⑤50°C孵育10min，如果酶切效果不佳，可延长至15min，加热时请尽量使用薄壁管。
2.PNGase F酶切-还原条件
① 用10×糖苷酶切反应液（500mM磷酸二氢钠，pH=7.5）将酶增效表面活性剂复溶至终浓度10mg/ml，轻柔混匀。
② 用去离子水溶解50-100μg糖蛋白，加入5μl含本品的10×糖苷酶切反应液，1M DTT至终浓度20mM（按总体积50μl计算），并用去离子水定溶至48-49μl。
③ 80-90°C加热3-5min，冷却至室温（此步骤不可省略）。
④ 加入1-2μl的PNGase F，使得反应总体积为50μl，混匀。
⑤ 50°C孵育10min，如果酶切效果不佳，可延长至15min，加热时请尽量使用薄壁管。
3.制备兼容LC和LC/MS的样品
①若需分析N-聚糖，可在反应结束后加入甲酸（终浓度0.5%）或TFA（终浓度0.5%）或HCl（终浓度30-50mM），于37°C孵育30-45min以降解酶增效表面活性剂 ，降解过程中会出现沉淀，可通过10000g离心10min去除沉淀，上清可直接用于后续的HPLC或质谱分析。
②若需分析去糖基化蛋白，可直接采用SDS-PAGE检测脱糖效果，也可通过微量透析或微过滤交换缓冲液后用于HPLC和质谱分析。
4.Trypsin酶切
① 用50mM碳酸氢铵溶液将酶增效表面活性剂复溶至终浓度10mg/ml，轻柔混匀。
②在蛋白溶液中加入含酶增效表面活性剂的50mM碳酸氢铵溶液至终浓度为1mg/ml，混匀。
③ 加入DTT溶液至终浓度5mM，60度加热30min，加热结束后冷却至室温。
④加入IAA至终浓度15mM，室温避光放置15min。
⑤加入Trypsin进行酶切，添加比例建议1:50-1:100，37°C孵育3h-过夜。
⑥反应结束后加入甲酸（终浓度0.5%）或TFA（终浓度0.5%）或HCl（终浓度30-50mM），于37°C孵育30-45min以降解酶增效表面活性剂 ，降解过程中会出现沉淀，可通过10000g离心10min去除沉淀，上清可直接用于后续的HPLC或质谱分析。
保存方法
本品应存放于-20°C，可保存至少24个月；复溶后可在-80°C保存1个月，2-8°C保存1周。
Enzyme-Enhancing Surfactant is an anionic, mild protein denaturant specifically designed to increase the hydrolysis efficiency of proteases (such as trypsin, Lys‑C, PNGase F, Asp‑N, etc.) in solution. By promoting protein solubilization and structural unfolding, it allows endoproteases to more readily access cleavage sites within hydrophobic proteins, thereby significantly improving digestion efficiency and substantially shortening digestion time.
Unlike conventional denaturants, this surfactant has virtually no adverse effect on the activity of the above‑mentioned hydrolytic enzymes at final concentrations below 1%. At the same time, it avoids introducing unwanted modifications such as oxidation or deamidation to the target proteins and is fully compatible with mass spectrometry detection.
Owing to its mild yet effective denaturing properties, it also improves the solubility of hydrophobic proteins and peptides, reduces the amount of costly protease required, and helps laboratories complete high‑recovery digestion workflows in less time, meeting the demands of high‑throughput analysis.
Product Applications
Enhance the cleavage efficiency of proteolytic enzymes and shorten digestion time
Instructions (For Reference Only)
Before use, centrifuge the product at 10,000 × g for 10 s (applicable only to the 1 mg and 3 mg sizes) to ensure all powder settles at the bottom of the tube.
1.PNGase F Digestion – Non‑Reducing Conditions
① Reconstitute the Enzyme-Enhancing Surfactant in 10× Glycosidase Digestion Buffer (500 mM sodium dihydrogen phosphate, pH 7.5) to a final concentration of 10 mg/mL. Mix gently.
② Dissolve 50–100 μg of glycoprotein in deionized water. Add 5 μL of the 10× Glycosidase Digestion Buffer containing the surfactant, and bring the volume to 48–49 μL with deionized water.
③ Heat at 80–90 °C for 3–5 min, then cool to room temperature (this step is optional, but heating improves digestion efficiency).
④ Add 1–2 μL of PNGase F to bring the total reaction volume to 50 μL. Mix well.
⑤ Incubate at 50 °C for 10 min. If digestion is suboptimal, extend to 15 min. Use thin‑walled tubes whenever possible during heating.
2.PNGase F Digestion – Reducing Conditions
① Reconstitute the Enzyme-Enhancing Surfactant in 10× Glycosidase Digestion Buffer (500 mM sodium dihydrogen phosphate, pH 7.5) to a final concentration of 10 mg/mL. Mix gently.
② Dissolve 50–100 μg of glycoprotein in deionized water. Add 5 μL of the 10× Glycosidase Digestion Buffer containing the surfactant and 1 M DTT to a final concentration of 20 mM (based on a total volume of 50 μL), then bring the volume to 48–49 μL with deionized water.
③ Heat at 80–90 °C for 3–5 min, then cool to room temperature (this step is essential and cannot be omitted).
④ Add 1–2 μL of PNGase F to bring the total reaction volume to 50 μL. Mix well.
⑤ Incubate at 50 °C for 10 min. If digestion is suboptimal, extend to 15 min. Use thin‑walled tubes whenever possible during heating.
3.Preparation of Samples Compatible with LC and LC/MS
① For N‑glycan analysis, after the digestion is complete, add formic acid (final concentration 0.5%), TFA (final concentration 0.5%), or HCl (final concentration 30–50 mM). Incubate at 37 °C for 30–45 min to degrade the Enzyme-Enhancing Surfactant. Precipitation may occur during degradation. Remove the precipitate by centrifugation at 10,000 × g for 10 min. The supernatant can be directly used for subsequent HPLC or mass spectrometry analysis.
② For deglycosylated protein analysis, the deglycosylation efficiency can be assessed directly by SDS‑PAGE. Alternatively, after buffer exchange via micro‑dialysis or microfiltration, the sample can be used for HPLC and mass spectrometry analysis.
4.Trypsin Digestion
① Reconstitute the Enzyme-Enhancing Surfactant in 50 mM ammonium bicarbonate solution to a final concentration of 10 mg/mL. Mix gently.
② Add the reconstituted surfactant solution (in 50 mM ammonium bicarbonate) to the protein solution to a final surfactant concentration of 1 mg/mL. Mix well.
③ Add DTT solution to a final concentration of 5 mM. Heat at 60 °C for 30 min, then cool to room temperature.
④ Add IAA to a final concentration of 15 mM and incubate in the dark at room temperature for 15 min.
⑤ Add trypsin for digestion (recommended enzyme‑to‑substrate ratio 1:50–1:100) and incubate at 37 °C for 3 h–overnight.
⑥ After digestion, add formic acid (final concentration 0.5%), TFA (final concentration 0.5%), or HCl (final concentration 30–50 mM). Incubate at 37 °C for 30–45 min to degrade the Enzyme-Enhancing Surfactant. Precipitation may occur during degradation. Remove the precipitate by centrifugation at 10,000 × g for 10 min. The supernatant can be directly used for subsequent HPLC or mass spectrometry analysis.
Storage Conditions
Store at –20 °C; stable for at least 24 months. After reconstitution, store at –80 °C for up to 1 month or at 2–8 °C for up to 1 week.
规格
规格或纯度
BioReagent, ≥95%(NMR)
稳定性与储存
Store at -20℃ long term (24 months).
英文名称
Enzyme Boost Surfactant
储存条件
-20°C储存
运输条件
超低温运输
纯度
≥95%(NMR)
产品线
生物产品
技术文档
高级数据
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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 |
|---|---|---|
 ZJ26F0939343 |
分析证书 | B1526913 |
| ZJ26F0939342 |
分析证书 | B1526913 |
| ZJ26F0939341 |
分析证书 | B1526913 |
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