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應用
胰蛋白酶消化多肽时,胰蛋白酶:多肽使用约 1:100 至 1:20 的比例。该产品的典型用途是去除培养表面的贴壁细胞。使细胞脱离其底物所需的胰蛋白酶浓度主要取决于细胞类型和培养时间。胰蛋白酶也被用于细胞培养过程中的细胞重悬,蛋白质组学研究中的蛋白质消化和各种凝胶消化。其他的应用还包括通过基于膜的技术评估结晶,并在一项研究中确定蛋白质折叠速率和产量可能受到动力学陷阱存在的限制。
生化/生理作用
胰蛋白酶切割赖氨酸和精氨酸残基 C 末端的肽。如果酸性残基位于裂解位点两侧,则该反应的水解速率减慢,如果脯氨酸残基位于裂解位点的羧基侧,则水解停止。胰蛋白酶活性的最适 pH 为 7-9。胰蛋白酶也可裂解氨基酸合成衍生物的酯键和酰胺键。向胰蛋白酶溶液中加入 EDTA,作为螯合剂,可中和钙和镁离子,使胰蛋白酶作用的肽键模糊。去除这些离子会增加酶的活性。
DFP、TLCK、APMSF、AEBSEF和抑肽酶等丝氨酸蛋白酶抑制剂可抑制胰蛋白酶。
DFP、TLCK、APMSF、AEBSEF和抑肽酶等丝氨酸蛋白酶抑制剂可抑制胰蛋白酶。
注意
本品应在-20℃ 冷冻保存。
單位定義
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。
準備報告
此溶液为25 g/L猪胰蛋白酶的Hank平衡盐溶液(含酚红)。
訊號詞
Warning
危險聲明
危險分類
Skin Sens. 1
儲存類別代碼
10 - Combustible liquids
水污染物質分類(WGK)
WGK 3
閃點(°F)
Not applicable
閃點(°C)
Not applicable
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实验方案
胰蛋白酶可用于从培养表面去除贴壁细胞。从培养基底物除去细胞的最常用方法是用胰蛋白酶或胰蛋白酶/EDTA溶液处理。
胰蛋白酶常用于从表面进行贴壁细胞的解离。我们可提供一系列不同的胰蛋白酶溶液来满足您的特定细胞系需求。
Trypsin is commonly used for dissociating adherent cells from surfaces. A wide variety of trypsin solutions are available to meet your specific cell line requirements.
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