转谷氨酰胺酶活性检测试剂盒 比色法、Transglutaminase Activity Assay Kit, Colorimetric详细如下: 转谷氨酰胺酶活性检测试剂盒为高通量筛选 TG 调节剂和抑制剂提供了一种方便的检测方法。TG 催化聚-D-赖氨酸的游离胺基团与生物素-TVQQEL 肽底物中谷氨酰胺的γ-羧酰胺基团之间形成共价键,该胺基团包被在板表面。该反应将生物素缀合的肽固定在板表面。使用链霉亲和素-辣根过氧化物酶 (HRP) 和 TMB 底物测定固定化生物素的量。HRP 和 TMB 底物产生的比色信号与样品中 TG 的活性成正比。测定以方便的 96 孔微孔板形式进行。 转谷氨酰胺酶 (TG) 是催化蛋白质翻译后修饰的 Ca2+ 和硫醇依赖性酶。这些共价反应包括在蛋白质结合的谷氨酰胺残基的羧酰胺基团和蛋白质结合的赖氨酸和多胺残基的ε-氨基之间形成异肽键,谷氨酰胺残基的水解和酯化。TG 家族至少包括九个成员:TG 1-7、凝血因子 XIIIa 和角质形成细胞膜结合的 TG 形式。TG作为潜在的药物靶点引起了相当大的兴趣。 1 Add TG enzyme diluent and test compounds into 96-well plate (Component A). Suggested volume for TG enzyme diluent is 40 uL/well and test compound is 10 uL/well. Establish the following controls at the same time: Positive control contains TG enzyme diluent without test compound. Inhibitor control contains TG enzyme diluent and a known TG inhibitor (such as Component J). Vehicle control contains TG enzyme diluent and vehicle used to deliver test compound (e.g. DMSO). Test compound control contains reaction buffer and test compound. Substrate control contains reaction buffer only. Note: Bring the total volume of all the controls to 50 uL for 96-well plate with reaction buffer.