蛋白芯片
信号通路磷酸化广筛抗体芯片（PEX100）
从细胞膜、胞浆到细胞核，存在多条信号通路串联交叉形成的复杂信号网络。该信号网络在细胞受到胞外刺激后将信号通过级联放大、分散调节等方式传入胞内，引起一系列的综合性细胞应答。一种生物效应的出现往往存在多条信号通路的同步活化，可逆的磷酸化修饰反应则是细胞内部最为普遍和节能的信号蛋白活化调节方式。因此，找到激活的信号通路乃至发生磷酸化调变的通路蛋白，往往成生命科学研究的起点。
信号通路磷酸化广谱筛选抗体芯片（PEX100），采用三维高分子膜专用技术，在片基上高密度结合 1318 种高特异抗体，分别检测432个信号蛋白的679个磷酸化位点。这些信号蛋白广泛参与多条重要信号通路信号传导过程。芯片针对每一个特定蛋白磷酸化位点，设置一对抗体分别检测其磷酸化（Phospho）和非磷酸化（non-Phospho）状态以提高磷酸化检测灵敏度和稳定性。一次芯片实验即可实现30多条信号通路的同步筛选和具体调变位点的清晰定位，为后续生物现象的深入探索提供明确的研究方向。
| 抗体芯片特点
● 芯片规格为76 x 25 x 1 mm；
● 实现31条信号通路全面筛选；
● 每种抗体设置2次技术重复；
● 适用于组织、细胞等多类型样本；
● 5 x106细胞、100μg总蛋白量即可满足实验；
● 每个检测位点设有磷酸化和非磷酸化配对抗体；
● 可通用于人、小鼠、大鼠等多类型模式生物检测。
| 芯片检测范围
|
信号通路 |
蛋白数 |
信号通路 |
蛋白数 |
|---|---|---|---|
|
MAPK signaling pathway |
67 |
Interleukin signaling pathway | 37 |
|
Erk signaling pathway |
59 |
Regulation of actin cytoskeleton |
36 |
|
Angiogenesis |
57 |
Jak-STAT signaling pathway |
32 |
|
Focal adhesion |
56 |
FGF signaling pathway |
29 |
|
Neurotrophin signling pathway |
52 |
Endocytosis |
29 |
|
ErbB signaling pathway |
45 |
Integrin signaling pathway |
29 |
|
AKT signaling pathway |
45 |
VEGF signaling pathway |
27 |
|
Chemokine signaling pathway |
45 |
Wnt signaling pathway |
26 |
|
PDGF signaling pathway |
43 |
p53 signaling pathway |
25 |
|
T cell receptor signaling pathway |
43 |
NF-kappa B signaling pathway | 25 |
|
Apoptosis signaling pathway |
41 |
Adipocytokine signaling pathway |
19 |
|
EGF receptor signaling pathway |
41 |
Calcium signaling pathway |
19 |
|
Insulin signaling pathway |
39 |
Insulin/IGF pathway |
16 |
|
CREB signaling pathway |
39 |
mTOR signaling pathway |
16 |
|
Cell cycle |
37 |
TGF-beta signaling pathway |
13 |
| 抗体芯片原理
| 客户案例
PEX100芯片协助揭秘衰老引起的营养不良潜在机制
研究人员采集了不同月龄的正常B6小鼠的小肠进行组织学观察寻找到衰老引起营养吸收障碍的靶器官主要在空肠绒毛隐窝区域。经文献调研及实验研究发现，mTORC1蛋白在老龄小鼠肠绒毛老化过程中发挥着关键作用，但mTORC1通路是如何调控肠绒毛老化的呢？研究人员果断采用PEX100磷酸化广筛抗体芯片技术，对Villin-Cre;Tsc1f/f小鼠的小肠组织进行广泛的蛋白磷酸化筛选。抗体芯片迅速锁定了mTORC1下游一系列关键分子，其中p38MAPK蛋白的在Villin-Cre;Tsc1f/f小鼠中上调7.21倍，尤为显著。
研究人员利用Westernblot证实了p38MAPK以及其上游的MKK6蛋白在Tsc1−/−肠绒毛中显著上调，而雷帕霉素的施用可以逆转该现象。为进一步证实MKK6与p38MAPK的调控关系，在体外培养的MEFs细胞中，利用顺转siRNA的方式敲降MKK6后，发现p38MAPK的磷酸化变化受到了MKK6的调控，即说明MKK6是介于mTORC1和p38MAPK的中间信号转导蛋白。
研究人员通过一系列的研究，逐步明确了mTORC1-MKK6-p38MAPK-p53通路在肠绒毛老化中发挥着关键作用，并为逆转肠绒毛老化，改善衰老引起的营养不良找到了潜在的治疗靶点和候选药物小分子。
详细内容点击查看：https://mp.weixin.qq.com/s/_tG9Y9v-5oh3qPDvAtPGjw
| 客户文献
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