我的实验是用VEGF处理细胞后测定细胞PKC激活水平,拟采用Western blot的方法,用ECL法检测,这样分析PKC激活程度结果国际上认可吗?
我在Cell Signaling Technology上,查到
2261 Phospho-(Ser) PKC Substrate Antibody W IP E D
9371 Phospho-PKC (pan) (betaII Ser660) Antibody W IP
9921 Phospho-PKC Antibody Sampler Kit
9375 Phospho-PKCalpha/beta II (Thr638/641) Antibody W IP
9374 Phospho-PKCdelta (Thr505) Antibody W IHC
9376 Phospho-PKCdelta/theta (Ser643/676) Antibody W IHC
9377 Phospho-PKCtheta (Thr538) Antibody W IHC
9378 Phospho-PKCzeta/lambda (Thr410/403) Antibody W IHC
我只想要检测PKC的激活程度,并不要求激活哪种亚型,应该用那一种呢?
licanming wrote: 我的实验是用VEGF处理细胞后测定细胞PKC激活水平,拟采用Western blot的方法,用ECL法检测,这样分析PKC激活程度结果国际上认可吗?
我在Cell Signaling Technology上,查到
2261 Phospho-(Ser) PKC Substrate Antibody W IP E D
9371 Phospho-PKC (pan) (betaII Ser660) Antibody W IP
9921 Phospho-PKC Antibody Sampler Kit
9375 Phospho-PKCalpha/beta II (Thr638/641) Antibody W IP
9374 Phospho-PKCdelta (Thr505) Antibody W IHC
9376 Phospho-PKCdelta/theta (Ser643/676) Antibody W IHC
9377 Phospho-PKCtheta (Thr538) Antibody W IHC
9378 Phospho-PKCzeta/lambda (Thr410/403) Antibody W IHC
我只想要检测PKC的激活程度,并不要求激活哪种亚型,应该用那一种呢?
楼上的说法很正确!PKC肯定不是广谱的被激活,但因为我暂未看到文献明确提示我模型中的通路究竟涉及哪个亚型,而我又没有那么多经费一个一个亚型去试,故我想能不能就检测总的PKC激活水平,毕竟有亚型被激活,那总的活化PKC水平也应该会升高啊!虽然敏感性和特异性肯定没有单独检测一个亚型的高。
还想请教:Phospho-PKC (pan) (gamma Thr514) Antibody 和Phospho-PKC (pan) (βII Ser660) Antibody 这两个抗体的针对的亚型有什么不同?既然都是pan,那是否说都是针对所有的亚型,只是磷酸化的残基不同?我在站里搜了一下pan的解释(来自rickerzhang):
Most protein families have different members which might share the common motif or sequence but have different function. Pan antibody can recognize the entire members in that family. It is always a good positive control for the specific member of one family.
For example:
1. Cadherins constitute a family of proteins, the members of which are differentially expressed in the different tissues. Cadherins are divided into type I and type II subgroups.
Type I cadherins include epithelial cadherin (E-cadherin, cadherin-1 or uvomorulin), neural cadherin (N-cadherin or cadherin-2), placental cadherin (P-cadherin or cadherin-3) and retinal cadherin (R-cadherin or cadherin-4). Kidney cadherin (K-cadherin or cadherin-6) and osteoblast cadherin (OB-cadherin or cadherin-11) are type II cadherins.
Pan Cadherin antibody can reacts with ALL cadherins, it is a good Plasma Membrane Marker.
2. One of the best characterized members is E-cadherin, which is prevalent in epithelial tissues. It has been shown to play a crucial role in the process of tumor cell metastasis.
If you want to detect E-cadherin only, you should use E-cadherin antibody instead of Pan Cadherin antibody
按他(她)这样说应该一个带pan的抗体就可以检测所有PKC亚型了,那为什么还要分磷酸化不同的残基呢?
谢谢!