Phospho-Akt (Thr450) Antibody

【求助】磷酸化的Akt不见了！！

米宝 这俩天做实验，极其郁闷。我做的总AKt和pAkt,大鼠脑组织，第一次跑总AKT的时候，我觉得可能是因为蛋白是提取的浓度比较好吧，出来了一条浓浓的条带，三组总AKt都出了，接下来做pAkt,就出了一个带，效果挺明显的。后来再做pAkt的时候，跑了4张膜，显影的时候，太干净了，上面什么都没有，甚至连个杂带，斑点 都没！实验是在实验员的领导下完成的，技术上基本没什么纰漏，查阅本版面有关这问题方面的介绍，有的解释说pakt降解了！一抗是用的Phospho-Akt

Using Phospho‐Motif Antibodies to Determine Kinase Substrates

comprising both the phosphorylated residue and the surrounding residues that determine kinase specificity, with degenerate residues taking up the remaining positions. Currently, several categories of phospho?motif antibody are commercially available

Optimized Protocol to Make Phospho-Specific Antibodies that Work

, not simply its level of expression. In this review, we will discuss both the design of the phosphopeptide immunogen and immunization. The affinity purification of the phospho-specific antibody as well as the methods most suitable for characterizing