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各2ml/各10ml
| 规格: | 各2ml | 产品价格: | ¥358.0 |
|---|---|---|---|
| 规格: | 各10ml | 产品价格: | ¥1429.0 |
| 产品编号 | 产品名称 | 产品包装 | 产品价格 |
| 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 | 358.00元 |
| 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 | 1429.00元 |
碧云天的蛋白酶磷酸酶抑制剂混合物(哺乳动物样品抽提用, 50X) (Protease and phosphatase inhibitor cocktail for mammalian cell and tissue extracts, 50X) 是一种用于哺乳动物细胞或组织蛋白提取的蛋白酶和磷酸酶抑制剂混合物组合。本品含有一管蛋白酶抑制剂(哺乳动物样品抽提用, 50X) (52.5mM AEBSF, 40μM Aprotinin, 2.5mM Bestatin, 0.75mM E64, 1mM Leupeptin and 0.75mM Pepstatin A in DMSO) 以及一管磷酸酶抑制剂(50X) (250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate and 50mM sodium orthovanadate in H2O ),并提供独立包装的0.25M EDTA。
哺乳动物细胞或组织提取物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哺乳动物细胞或组织蛋白提取的蛋白酶和磷酸酶抑制剂混合物,包含了广谱的丝氨酸、半胱氨酸和酸性蛋白酶抑制剂/氨基肽酶抑制剂,以及丝氨酸/苏氨酸、酪氨酸、酸性及碱性磷酸酶抑制剂。
本蛋白酶磷酸酶抑制剂混合物含52.5mM AEBSF, 40μM Aprotinin, 2.5mM Bestatin, 0.75mM E64, 1mM Leupeptin, 0.75mM Pepstatin A, 250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate, 50mM sodium orthovanadate, 分别配制在高纯度DMSO和超纯水中。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1:50的比例分别把蛋白酶抑制剂混合物(哺乳动物样品抽提用, 50X),磷酸酶抑制剂(50X)和0.25M EDTA加入裂解液中,即可用于哺乳动物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哺乳动物细胞或组织提取物中的各种蛋白酶(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)和磷酸酶(如丝氨酸/苏氨酸、酪氨酸、酸性及碱性磷酸酶等)。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;Pepstatin A是天冬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;Sodium fluoride是酸性磷酸酶可逆抑制剂;焦磷酸钠(sodium pyrophosphate)是丝氨酸/苏氨酸磷酸酶不可逆抑制剂;β-甘油磷酸(β-glycerophosphate)是丝氨酸/苏氨酸磷酸酶可逆抑制剂;正钒酸钠(sodium orthovanadate)是碱性磷酸酶和酪氨酸磷酸酶可逆抑制剂。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: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 |
| P1050-1 | 蛋白酶抑制剂混合物(哺乳动物样品抽提用, 50X) | 2ml |
| P1050-2 | 磷酸酶抑制剂混合物(50X) | 2ml |
| P1050-3 | 0.25M EDTA, pH 8.0 | 2ml |
| — | 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和磷酸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。
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哺乳动物组织样品RNA的抽提最基本的要素是防止RNA在抽提过程中降解,以及尽可能得到最高的RNA抽提效率。以下介绍我们实验室认为最符合以上两个要素的抽提方法。一、实验材料:Trizol试剂(Invitrogen公司)研钵,匀浆器,镊子,铝箔都高温灭菌即可;二、具体过程:1、样品在离体后应迅速冷冻在液氮中,若是比较大块的组织,要尽量把组织剪切成黄豆大小的块,使液氮迅速渗透到组织内部而防止组织内部的RNA降解。组织用铝箔包好或放入冻存管中,并做好记号,注意不要把记号写在纸上,以防纸被组织的血水渗透
相关专题 蛋白酶抑制剂 破碎细胞提取蛋白质的同时可释放出蛋白酶,这些蛋白酶需要迅速的被抑制以保持蛋白质不被降解。在蛋白质提取过程中,需要加入蛋白酶抑制剂以防止蛋白水解。以下列举了5种常用的蛋白酶抑制剂和他们各自的作用特点,因为各种蛋白酶对不同蛋白质的敏感性各不相同,因此需要调整各种蛋白酶的浓度。由于蛋白酶抑制剂在液体中的溶解度极低,尤其应注意在缓冲液中加人蛋白酶抑制剂时应充分混匀以减少蛋白酶抑制剂的沉淀。在宝灵曼公司
剂的蛋白质抽提剂(107 个细胞中加入1ml抽提试剂;5×106 个细胞中加入0.5ml抽提试剂),轻轻摇动5分钟5、细胞悬浮液到离心管中,冰浴下摇动15分钟进行裂解。6、裂解液于预冷的离心机中14,000×g离心15分钟。弃去沉淀,上清液蛋白质抽提物立刻转移入新的离心管中保存待用(-80o C或液氮中)。三、哺乳动物组织的蛋白质抽提步骤1、组织称重,切小块放入管中。3、PriCells蛋白质抽提剂(1ml抽提试剂中加入5μl蛋白酶抑制剂混合液\5μlPMSF和5μl磷酸酶混合液等等)。3、加入预冷
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