计算溶液所需的质量、体积或浓度。
ActiBioPure™,EnzymoPure™,His标签,生物活性,质谱级 (MS),重组,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。
2-8°C储存。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。
V8蛋白酶属于丝氨酸蛋白酶家族,能特异水解谷氨酸(Glutamic acid、E)或天冬氨酸(Aspartic acid、D)残基羧基侧肽键。在pH 7.8的NH₄HCO₃和pH 4.0的 CH₃COONH₄缓冲液中识别并切割Glu羧基端肽键,在pH 7.8的磷酸盐缓冲液中识别并切割Glu或Asp羧基端肽键,且对Glu的水解速率高于Asp。
产品应用
在进行蛋白质酶切测序、肽图谱分析和肽质量指纹图谱分析中使用时,可以单独使用或与其他蛋白酶混合使用。
基本特性
V8蛋白酶在pH4.0-10.0之间均有活性,最适pH为8.0-8.5。V8蛋白酶的抑制剂有二异丙基氟磷酸(DFP)、α2-巨球蛋白和Nα-P-甲苯磺酰基-L-赖氨酸氯甲基酮(TLCK)。
使用方法
使用前注意:可按使用量分装以最大限度避免污染或自切。
目的蛋白溶解:用酶切buffer溶解目的蛋白,例如25-50 mM NH4HCO3 pH 7.8,若溶解度不好对目的蛋白变性,添加尿素、SDS、DTT或加热。尿素和SDS对V8蛋白酶的影响见表一。
酶切:V8蛋白酶和目的蛋白推荐比例为1/20-1/100(W/W),25℃或37℃酶切2-18h。
表一:V8蛋白酶在含有不同浓度变性剂,尿素25℃2h,SDS25℃12h的稳定性
(注:V8蛋白酶0.5mg/ml,50mM Tris-HCl pH8.0)

应用举例:重组V8酶切胰岛素
100 μl胰岛素溶液+198 μl 0.4 M Tris-HCl pH 8.0+60 μl 0.1%V8蛋白酶液+超纯水补至400 μl。37 ℃水浴反应2 h,加6 μl磷酸溶液终止反应,C18柱分析:

储存
储存稳定性:重组V8蛋白酶冻干粉存于2-8℃,24个月稳定;
溶解后稳定性:用水或50 mM Tris-HCl pH 8.0溶解以后存于-20℃ 以下(1mg/ml),6个月稳定;用水或50 mM Tris-HCl pH 8.0溶解以后存于4℃(1mg/ml,无菌),2个月稳定;用50 mM Tris-HCl pH 8.0溶解以后存于25℃或37℃(0.5mg/ml)12h,活性可维持85%以上;溶解后溶液可反复冻融10次无活性损失。
产品优势
序列分析纯胰蛋白酶:特异性高,稳定性好。
无动物源性:重组生产,无外源性的病毒污染,生产过程不使用任何动物源原料。
质量稳定:批量生产,可保证稳定连续的批次生产;产品批次间无差异,质量稳定。
纯度高:比活高;宿主蛋白残留小于生物制品限度要求。
冻干粉:易于储存和运输。
符合法规要求:生产设备和生产环境符合相关法规要求,生产过程完全遵循ISO 9001:2015质量体系并符合GMP指导原则。
质量文件完整:按客户需求,可提供相关法规支持文件。
V8 protease belongs to the serine protease family and specifically hydrolyzes the peptide bonds on the carboxyl side of glutamic acid (Glu, E) or aspartic acid (Asp, D) residues. It recognizes and cleaves the carboxyl-terminal peptide bonds of Glu in NH₄HCO₃ buffer (pH 7.8) and CH₃COONH₄ buffer (pH 4.0). In phosphate buffer (pH 7.8), it recognizes and cleaves the carboxyl-terminal peptide bonds of both Glu and Asp, with a higher hydrolysis rate for Glu than for Asp.
Product Applications
Basic Properties
V8 protease is active in the pH range of 4.0–10.0, with an optimal pH of 8.0–8.5.
Inhibitors of V8 protease include diisopropyl fluorophosphate (DFP), α₂-macroglobulin, and Nα-p-tosyl-L-lysine chloromethyl ketone (TLCK).
Usage Method
Precautions before use: Aliquot according to the required amount to minimize contamination or autolysis.
Dissolution of target protein: Dissolve the target protein in digestion buffer (e.g., 25–50 mM NH₄HCO₃, pH 7.8). If solubility is poor, denature the target protein by adding urea, SDS, DTT, or heating. The effects of urea and SDS on V8 protease are shown in Table 1.
Digestion: The recommended mass ratio of V8 protease to target protein is 1:20–1:100 (W/W). Perform digestion at 25°C or 37°C for 2–18 hours.


Storage
Storage stability: Lyophilized recombinant V8 protease is stable for 24 months when stored at 2–8°C.
Product Advantages
Sequence-grade trypsin (analytical pure): High specificity and good stability.
Animal-origin-free: Produced by recombination, no exogenous virus contamination, and no animal-derived raw materials used in the production process.
Stable quality: Manufactured in batches to ensure stable and continuous production; no variation between batches, with consistent quality.
High purity: High specific activity; residual host protein is below the limit required for biological products.
Lyophilized powder: Easy to store and transport.
Compliance with regulatory requirements: Production equipment and environment meet relevant regulatory requirements; the production process fully complies with the NSF ISO 9001:2015 quality system and GMP guidelines.
Complete quality documents: Relevant regulatory support documents can be provided upon customer request.
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 | E1505824 | |
| 分析证书 | E1505824 |
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 Recombinant 级别指南 → 查看 Suitable for mass spectrometry (MS) 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 →