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文献和实验运用Cell Based Elisa检测信号通路蛋白和磷酸化蛋白
using Phospho-p38 and Total-p38 antibodies (C). Cell-Based Elisa将小鼠巨噬细胞4/4细胞种在96孔板内,每个孔种5 x 104 cells/cm2细胞。细胞用不同浓度的anisomycin刺激,我们来监测P38 MAPK和JNK两种蛋白的变化。按照试剂盒的实验步骤,分别固定,打孔,猝灭,封闭,然后一抗孵育,二抗孵育,最后显色,用常规的酶标仪读取数值。最后数据再和试剂盒提供的结晶紫核染料结果做纠正,最后得出以上数据。Quantitative
This protocol is based on enrichment from 1 x 107 leucocytes (MNC). It is scalable from 1 x 107 –2 x 109 cells. Keep the cells and buffers cold (2–8°C) during the whole process. 1) Transfer 100 μl (1 x 107 ) leucocytes in Buffer
Using Phospho‐Motif Antibodies to Determine Kinase Substrates
in many signaling pathways typically associated with human malignancies. Phospho?motif antibodies are engineered to recognize proteins that contain a phosphorylated residue in the context of a specific motif. They are raised against a library of phospho?peptides
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