
Trypsin, TPCK Treated(胰蛋白酶(TPCK处理))
货号:LS003740
品名:Trypsin, TPCK Treated(胰蛋白酶(TPCK处理))
品牌:Worthington-biochem
规格:100mg
价格:请询价
货期:现货
关键词:
TPCK处理胰蛋白酶、Worthington生物化学、胰蛋白酶说明书、细胞培养用胰酶、蛋白质组学试剂、病毒扩增酶、质谱分析用酶、蛋白水解酶、科研级酶制剂、牛胰蛋白酶、TPCK修饰酶、实验室常用酶、高纯度胰酶、无外泌蛋白酶、低自溶胰酶、流感病毒研究试剂、MDCK细胞消化酶、Edman降解用酶、SDS-PAGE样品制备酶、重组蛋白酶
产品详情介绍
产品描述
货号:LS003740
英文名:Trypsin, TPCK Treated
中文名:胰蛋白酶(TPCK处理)
品牌:Worthington-Biochem
规格:100 mg
CAS号:9002-07-7
应用领域:蛋白质组学、细胞培养、病毒研究、蛋白质测序
产品特点
1. 高纯度与稳定性
通过色谱纯化技术去除杂蛋白及天然抑制剂,确保酶活性稳定。经TPCK化学修饰后,显著抑制糜蛋白酶(Chymotrypsin)活性,减少自溶现象。
2. 低自溶性与高特异性
修饰后的胰蛋白酶在37°C下自溶率低于0.1%,适用于长时间反应。特异性水解赖氨酸(Lys)和精氨酸(Arg)羧基端的肽键,避免非特异性切割。
3. 无动物源性污染
采用重组技术或严格筛选的动物源原料,确保病毒、支原体及外泌蛋白酶的零污染,符合生物安全标准。
4. 广泛兼容性
适用于哺乳动物、植物及微生物蛋白的酶解,支持SDS-PAGE、质谱分析及Edman降解等下游实验。
储存条件
• 短期储存:2–8°C避光保存,避免反复冻融。
• 长期储存:-20°C冻存,建议分装后保存以减少活性损失。
• 保质期:未开封状态下24个月(2–8°C),开封后建议6个月内用完。
工作原理
胰蛋白酶是一种丝氨酸蛋白酶,通过催化肽键水解实现蛋白质的特异性切割。TPCK处理通过共价修饰酶活性位点,不可逆抑制糜蛋白酶活性,同时保留胰蛋白酶的核心功能。其作用机制如下:
1. 底物识别:结合Lys或Arg残基的羧基端,形成酶-底物复合物。
2. 催化水解:水分子攻击肽键羰基碳,生成酰胺键断裂产物。
3. 抑制副反应:TPCK阻断糜蛋白酶对芳香族氨基酸的切割,提升实验特异性。
解决实验中的关键问题
1. 细胞培养中的细胞分离
用于胰酶消化法分离贴壁细胞,避免传统胰酶中糜蛋白酶对细胞膜的损伤。
2. 蛋白质组学中的自溶干扰
减少内源性糜蛋白酶引起的肽段污染,提高质谱鉴定的准确性。
3. 病毒培养中的酶需求
在流感病毒扩增中,优化TPCK胰酶浓度以促进病毒吸附与复制。
4. 复杂样本的消化效率
适用于植物、微生物等难消化样本,通过调整浓度提升水解率。
使用方法
1. 溶解与稀释
• 称取100 mg粉末,溶于10 mL 1 mM HCl(pH 2.0–3.0),轻轻涡旋至溶解。
• 稀释至工作浓度,建议使用PBS或Tris缓冲液。
2. 细胞消化
• 加入含0.25% EDTA的胰酶溶液,37°C孵育1–5分钟。
• 中止反应:添加含血清培养基终止消化。
3. 蛋白质酶解
• 样本溶解于尿素缓冲液,还原烷基化处理。
• 加入TPCK胰酶,37°C孵育12–18小时。
4. 病毒扩增
• 在无血清培养基中添加1–2 μg/mL TPCK胰酶,接种病毒MOI 0.001–0.1,37°C孵育72小时。
常见问题与解决方案
Q1:酶活性低于预期
• 原因:储存温度不当或反复冻融。
• 解决:分装后-20°C保存,使用前复溶至1 mM HCl。
Q2:反应中出现自溶峰
• 原因:TPCK修饰不完全或缓冲液pH偏移。
• 解决:优化缓冲液pH至8.0–8.5,添加1 mM CaCl₂稳定酶活性。
Q3:细胞消化后存活率低
• 原因:胰酶浓度过高或孵育时间过长。
• 解决:使用0.25%胰酶+0.02% EDTA,孵育时间控制在5分钟内。
Q4:质谱检测到未切割的完整蛋白
• 原因:酶/底物比不足或消化时间过短。
• 解决:增加酶量至1:20,延长孵育至24小时。
免责声明
1. 本产品仅供科研使用,不可用于诊断、治疗或人体应用。
2. 实验结果可能因操作条件、样本特性或仪器差异而波动,建议用户自行优化实验方案。
3. 产品储存及运输中若发生泄漏或活性丧失,厂商不承担直接责任。
4. 本文内容基于品牌公开资料及行业常规信息整理,仅个人观点,具体以品牌信息文档为准。
关键词:TPCK处理胰蛋白酶、Worthington生物化学、胰蛋白酶说明书、细胞培养用胰酶、蛋白质组学试剂、病毒扩增酶、质谱分析用酶、蛋白水解酶、科研级酶制剂、牛胰蛋白酶、TPCK修饰酶、实验室常用酶、高纯度胰酶、无外泌蛋白酶、低自溶胰酶、流感病毒研究试剂、MDCK细胞消化酶、Edman降解用酶、SDS-PAGE样品制备酶、重组蛋白酶
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品名 |
规格 |
品牌 |
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LS004176,CAS:9001.12.1, |
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3 bx |
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|
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4ku |
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|
LS003741,CAS:9002.07.7, |
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500mg |
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|
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200 un |
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LS02111,CAS:42613.33.2, |
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LS006123,CAS:9001.78.9, |
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25mg |
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|
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5mg |
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|
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10 gm |
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|
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Deoxyribonucleic Acid, Calf Thymus |
1 gm |
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|
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Chymotrypsin, Alpha, 1X |
10 gm |
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|
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|
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|
LK003153,, |
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3bx |
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|
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250mg |
Worthington-biochem |
|
LS004154,CAS:9001.12.1, |
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1gm |
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|
LS004081,CAS:9001.78.9, |
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5 vi |
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|
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|
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|
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|
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|
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|
LS006365,CAS:39445.21.1, |
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|
LK003300,, |
Neonatal Cardiomyocyte Isolation System |
1 kt |
Worthington-biochem |
|
LS02123,CAS:9002.07.7, |
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1mg |
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|
LS002976,CAS:84195.52.8, |
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500mg |
Worthington-biochem |
|
LS01502,CAS:37278.25.4, |
Ribonuclease T2, Recombinant |
250ku |
Worthington-biochem |
|
LS002512,, |
Hexokinase, Lyophilized |
10 ku |
Worthington-biochem |
|
LS004206,CAS:9001.12.1, |
Collagenase, Type 3, Filtered |
50mg |
Worthington-biochem |
|
LS006308,CAS:9003.98.9, |
Amino Acid Oxidase, D- |
25mg |
Worthington-biochem |