agency
USP/NF, meets USP testing specifications
form
solid
specific activity
≥2,500 USP units/mg solid
mol wt
23.8 kDa
purified by
crystallization
solubility
H2O: soluble, saline: soluble
application(s)
diagnostic assay manufacturing
storage temp.
−20°C
Quality Level
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Application
对于胰蛋白酶消化肽,胰蛋白酶:肽 使用约为 1:100 至 1:20 的比例。从培养表面除去粘附细胞是本产品的一个典型应用。 从其底物中去除细胞所需的胰蛋白酶浓度主要取决于细胞类型和培养物的年龄。 胰蛋白酶也被用于细胞培养期间细胞的再悬浮、蛋白质组学研究中的蛋白质消化和各种凝胶内消化。 其他应用包括:通过基于膜的技术评估结晶,用在确定动力学陷阱的存在会限制蛋白质的折叠速率和产量的一项研究中。
Biochem/physiol Actions
胰蛋白酶切割赖氨酸和精氨酸残基的 C 端肽。 如果酸性残基位于切割位点的任一侧,则该反应的水解速度减慢,并且如果脯氨酸残基位于切割位点的羧基侧,则停止水解。 胰蛋白酶活性的最适 pH 为 7-9。 胰蛋白酶还可以起到裂解氨基酸合成衍生物的酯和酰胺键的作用。 将 EDTA 作为螯合剂添加到胰蛋白酶溶液中,用于中和钙离子和镁离子,这些离子遮蔽胰蛋白酶作用的肽键。 去除这些离子会增加酶活性。
丝氨酸蛋白酶抑制剂,包括 DFP、TLCK、APMSF、AEBSEF 和抑肽酶等,将抑制胰蛋白酶。
丝氨酸蛋白酶抑制剂,包括 DFP、TLCK、APMSF、AEBSEF 和抑肽酶等,将抑制胰蛋白酶。
Preparation Note
以1 mg/mL的浓度溶于1 mM HCl。
Other Notes
使用 BAEE 作为底物,一个 BAEE 单元,将在 pH 7.6,25℃ 下,每分钟产生 0.001 的 A253。
胰蛋白酶由223个氨基酸残基的单链多肽组成,其通过从胰蛋白酶原去除N-末端六肽而产生(在Lys-Ile肽键处被切割)。氨基酸序列通过6个二硫键交联。这是胰蛋白酶、β-胰蛋白酶的天然形式。β-胰蛋白酶可以在Lys-Ser残基处发生自裂解,并产生α -胰蛋白酶。胰蛋白酶是丝氨酸蛋白酶家族的成员。
Disclaimer
1mM HCl中的溶液以等分试样形式可在-20℃稳定储存1年。Ca2+的存在也可减少胰蛋白酶的自溶并保持其在溶液中的稳定性。胰蛋白酶还可在2.0M尿素、2.0M盐酸胍或0.1%(w / v)SDS中保留其大部分活性。
signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Respiratory system
存储类别
11 - Combustible Solids
ppe
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
法规信息
低风险生物材料
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