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多肽与蛋白> 蛋白样品制备> 蛋白酶抑制剂
蛋白酶抑制剂混合物(通用型, 100X)
产品编号: P1005
产品包装:1ml
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价格: ¥ 307.00
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产品编号 产品名称 产品包装 产品价格
P1005 蛋白酶抑制剂混合物(通用型, 100X) 1ml 307.00元
P1006 蛋白酶抑制剂混合物(通用型, 100X) 5ml 1220.00元

碧云天的蛋白酶抑制剂混合物(通用型, 100X) (Protease inhibitor cocktail for general use, 100X) 是一种通用型用于细胞或组织蛋白提取等的蛋白酶抑制剂混合物(200mM AEBSF, 30μM Aprotinin, 13mM Bestatin, 1.4mM E64 and 1mM Leupeptin in DMSO),并提供独立包装的0.1M EDTA。
细胞或组织提取物等样品中含有许多内源性的蛋白酶、磷酸酶等,容易导致提取物中的蛋白降解或去修饰,从而影响后续的蛋白检测。因此在提取物等样品中添加适当的蛋白酶、磷酸酶等抑制剂是防止蛋白降解和去修饰的有效方法。
本产品为经优化和测试的用于常规细胞或组织蛋白提取等的蛋白酶抑制剂混合物,包含了广谱的丝氨酸、半胱氨酸和酸性蛋白酶抑制剂、以及氨基肽酶抑制剂。具体组分与Sigma及Millipore同类产品的比较见表1。
表1. 碧云天蛋白酶抑制剂混合物(通用型, 100X)与Sigma及Millipore同类产品的比较:

品牌 Beyotime Sigma Millipore
产品编号 P1005/P1006 P2714/S8820 539131
产品形式 100X 如配制为100X 如配制为100X
AEBSF 200mM 200mM 50mM
Aprotinin 30μM 30μM 15μM
Bestatin 13mM 13mM -
E64 1.4mM 1.4mM 0.1mM
Leupeptin 1mM 0.1mM 0.1mM
EDTA 100mM 100mM 50mM

      本蛋白酶抑制剂混合物含200mM AEBSF, 30μM Aprotinin, 13mM Bestatin, 1.4mM E64和1mM Leupeptin (参考表1),配制在高纯度DMSO中。EDTA为单独包装,以便选择性使用。如用于检测金属蛋白酶活性,则不宜添加EDTA。
      本产品使用便捷,通常以1:100的比例分别把蛋白酶抑制剂混合物(通用型, 100X)和0.1M EDTA加入裂解液中,即可用于常规的细胞或组织蛋白的提取,并有效抑制蛋白降解。
      不同类型的蛋白提取物中含有不同类型的蛋白酶。例如,哺乳动物细胞提取物中通常含有高丰度的丝氨酸蛋白酶,细菌细胞提取物通常含有较多的丝氨酸蛋白酶和金属蛋白酶,哺乳动物组织提取物中主要含有丝氨酸、半胱氨酸和金属蛋白酶,植物提取物中常含有大量的丝氨酸和半胱氨酸蛋白酶,还有一些含有天冬氨酸蛋白酶。为此碧云天针对不同类型样品的蛋白提取,提供了不同类型的蛋白酶、磷酸酶等抑制剂混合物以供选择,具体请参考附表1。
      本产品可以有效抑制常规细胞或组织提取物中的各种蛋白酶活性,如丝氨酸蛋白酶、氨基肽酶、半胱氨酸蛋白酶、苏氨酸和天冬氨酸蛋白酶、金属蛋白酶等。其中AEBSF是一种水溶性的丝氨酸蛋白酶不可逆抑制剂,其抑制常数与PMSF和DFP的抑制常数相似,可以有效抑制胰蛋白酶(trypsin)、糜蛋白酶(chymotrypsin)、纤溶酶(plasmin)、激肽释放酶(kallikrein)和凝血酶(thrombin)等蛋白酶,作为PMSF和DFP的替代物,AEBSF的毒性更低、水溶性和水溶液稳定性更好。Aprotinin是丝氨酸蛋白酶的竞争性可逆抑制剂;Bestatin是氨基肽酶可逆抑制剂;E64是半胱氨酸蛋白酶不可逆抑制剂;Leupeptin是丝氨酸和半胱氨酸蛋白酶可逆抑制剂;EDTA作为螯合剂是金属蛋白酶的可逆抑制剂。各种抑制剂的具体信息请参考附表2。
      蛋白酶、磷酸酶和去乙酰化酶抑制剂混合物的比较和选择请见:https://www.beyotime.com/support/lysis-inhibitor cocktail.htm
      一个包装的本产品通常足够用于100ml裂解液的配制。
包装清单:

产品编号 产品名称 包装
P1005-1 蛋白酶抑制剂混合物(通用型, 100X) 1ml
P1005-2 0.1M EDTA, pH 8.0 1ml
说明书 1份

保存条件:
      -20℃保存,一年有效。
注意事项:
      本产品可以适当分装后保存,并应在临使用前添加到裂解液等适当溶液中。
      本产品提供了独立包装的EDTA,可以选择性使用。例如,后续用于检测提取液中金属蛋白酶活性,则不宜添加EDTA。
      本产品中含有的蛋白酶抑制剂对人体有害,操作时请小心,并注意有效防护以避免直接接触人体或吸入体内。
      本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
      为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1. 本蛋白酶抑制剂混合物为100X的储存液,使用时按照1:100的比例加入到裂解液中(例如,1ml裂解液中加入10μl蛋白酶抑制剂混合物),混匀后即可使用。含有蛋白酶抑制剂混合物的裂解液宜现用现配,不宜配制后冻存待后续使用。根据需要,0.1M的EDTA也按照1:100的比例加入到裂解液中。待所需的抑制剂添加完毕并混匀后,就可以开始进行常规细胞或组织的裂解和蛋白提取。

附表1. 碧云天提供的不同类型的蛋白提取用蛋白酶、磷酸酶、去乙酰化酶等抑制剂混合物:

产品编号 产品名称 包装
P1005 蛋白酶抑制剂混合物(通用型, 100X) 1ml
P1006 蛋白酶抑制剂混合物(通用型, 100X) 5ml
P1008 蛋白酶抑制剂混合物(通用型, 质谱兼容, 50X) 1ml
P1009 蛋白酶抑制剂混合物(通用型, 质谱兼容, 50X) 5ml
P1010 蛋白酶抑制剂混合物(哺乳动物样品抽提用, 100X) 1ml
P1011 蛋白酶抑制剂混合物(哺乳动物样品抽提用, 100X) 5ml
P1015 蛋白酶抑制剂混合物(植物样品抽提用, 100X) 1ml
P1016 蛋白酶抑制剂混合物(植物样品抽提用, 100X) 5ml
P1020 蛋白酶抑制剂混合物(真菌或酵母抽提用, 100X) 1ml
P1021 蛋白酶抑制剂混合物(真菌或酵母抽提用, 100X) 5ml
P1025 蛋白酶抑制剂混合物(细菌抽提用, 100X) 1ml
P1026 蛋白酶抑制剂混合物(细菌抽提用, 100X) 5ml
P1030 蛋白酶抑制剂混合物(His-Tag蛋白纯化用, 100X) 1ml
P1031 蛋白酶抑制剂混合物(His-Tag蛋白纯化用, 100X) 5ml
P1045 蛋白酶磷酸酶抑制剂混合物(通用型, 50X) 各2ml
P1046 蛋白酶磷酸酶抑制剂混合物(通用型, 50X) 各10ml
P1048 蛋白酶磷酸酶抑制剂混合物(通用型, 质谱兼容, 50X) 各1ml
P1049 蛋白酶磷酸酶抑制剂混合物(通用型, 质谱兼容, 50X) 各5ml
P1050 蛋白酶磷酸酶抑制剂混合物(哺乳动物样品抽提用, 50X) 各2ml
P1051 蛋白酶磷酸酶抑制剂混合物(哺乳动物样品抽提用, 50X) 各10ml
P1055 蛋白酶磷酸酶抑制剂混合物(植物样品抽提用, 50X) 各2ml
P1056 蛋白酶磷酸酶抑制剂混合物(植物样品抽提用, 50X) 各10ml
P1060 蛋白酶磷酸酶抑制剂混合物(真菌或酵母抽提用, 50X) 各2ml
P1061 蛋白酶磷酸酶抑制剂混合物(真菌或酵母抽提用, 50X) 各10ml
P1065 蛋白酶磷酸酶抑制剂混合物(细菌抽提用, 50X) 各2ml
P1066 蛋白酶磷酸酶抑制剂混合物(细菌抽提用, 50X) 各10ml
P1081 磷酸酶抑制剂混合物A (50X) 2ml
P1082 磷酸酶抑制剂混合物A (50X) 10ml
P1086 磷酸酶抑制剂混合物B (50X) 2ml
P1087 磷酸酶抑制剂混合物B (50X) 10ml
P1091 磷酸酶抑制剂混合物C (50X) 2ml
P1092 磷酸酶抑制剂混合物C (50X) 10ml
P1096 磷酸酶抑制剂混合物D (50X) 2ml
P1097 磷酸酶抑制剂混合物D (50X) 10ml
P1112 去乙酰化酶抑制剂混合物(100X) 1ml
P1113 去乙酰化酶抑制剂混合物(100X) 5ml

附表2. 常用蛋白酶、磷酸酶、去乙酰化酶抑制剂的具体相关信息:

名称 分子量 目标酶家族 抑制类型
AEBSF 239.5 丝氨酸蛋白酶 不可逆
Aprotinin 6511.5 丝氨酸蛋白酶 可逆
Bestatin 308.38 氨基肽酶 可逆
E64 357.4 半胱氨酸蛋白酶 不可逆
Leupeptin 475.6 丝氨酸和半胱氨酸蛋白酶 可逆
Pepstatin A 685.9 天冬氨酸蛋白酶 可逆
EDTA 372.24 金属蛋白酶 可逆
Phenanthroline 198.22 金属蛋白酶 可逆
Phosphoramidon 587.5 金属蛋白酶 可逆
Sodium fluoride 41.99 酸性磷酸酶 可逆
Sodium pyrophosphate 221.9 丝氨酸/苏氨酸磷酸酶 不可逆
β-glycerophosphate 306.11 丝氨酸/苏氨酸磷酸酶 可逆
Imidazole 68.2 碱性磷酸酶 可逆
Sodium molybdate 205.9 酸性磷酸酶 不可逆
Sodium tartrate dihydrate 230.1 酸性磷酸酶 可逆
Sodium orthovanadate 183.91 碱性和酪氨酸磷酸酶 可逆
(-)-p-Bromotetramisole oxalate 373.22 碱性磷酸酶 不可逆
Cantharidin 196.20 蛋白磷酸酶2A 可逆
Microcystin-LR 995.17 蛋白磷酸酶1和蛋白磷酸酶2A 可逆
Calyculin A 1009.17 蛋白磷酸酶1和蛋白磷酸酶2A 可逆
Trichostatin A 302.37 class I/II HDAC 可逆
EX-527 248.71 SIRT1 over SIRT2 & SIRT3 可逆
Nicotinamide 122.12 class III HDAC 可逆
Sodium butyrate 110.09 class I/II HDAC 可逆


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