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文献和实验secondary antibody were purchased from GIBCO-BRL. Phosphospecific rabbit polyclonal antibodies against 180Thr and 182Tyr dual-phosphorylated p38, against total p38 (phosphorylation state independent), against 204Tyr-phosphorylated ERK1/2 and against total
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
Using Phospho‐Motif Antibodies to Determine Kinase Substrates
, which may be used to specifically detect Ser, Thr, Ser/Thr, or Tyr residues phosphorylated by different protein kinase families. These antibodies are commonly used in immunoprecipitation and/or immunoblotting protocols to determine kinase?induced substrate
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