Overview:
CaM Kinase II (also known as CAMK2 and calcium/calmodulin-dependent protein kinase type II alpha chain) is a prominent kinase in the central nervous system that may function in the long-term potentiation of neurotransmitter release. CaM Kinase II autophosphorylates itself at Thr-286 which allows the kinase to switch from a calmodulin-dependent to a calmodulin-independent state. CaM Kinase II is composed of four different chains: alpha, beta, gamma, and delta. The different isoforms assemble into homo- or heteromultimeric holoenzymes composed of 8 to 12 subunits. This kinase is expressed in brain tissue. Alternative splicing occurs for this gene product.
References:
1. Lin,C.R., et al.: Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase. Proc. Natl. Acad. Sci. U.S.A. 1987: 84 (16); 5962-5966. 2. Bulleit,R.F., et al.: Conserved and variable regions in the subunits of brain type II Ca2+/calmodulin-dependent protein kinase. Neuron 1. 1988: (1); 63-72.Hanley,R.M., et al.: Functional analysis of a complementary DNA for the 50-kilodalton subunit of calmodulin kinase II. Science.1987: 237 (4812); 293-297.
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(1)确定抗体的名字,注意中英文名字、别名、亚型等信息。
(2)确定您的实验类型,Elisa,WB,IHC,ICC,或者是FACS。每种抗体说明书都会列出抗体经验证过适用的实验类型,如果抗体说明书没有提及的应用类型,并不意味着该抗体不适用于此种分析应用类型,而仅是说明尚未经过此种实验验证。
(3)确定实验样本的种属,Human,Mouse,Rat等。我们的抗体说明书都列出该抗体验证过的适用物种,可根据说明书列出已验证的种属来选择适合你实验的抗体。
(4)样本抗原蛋白的结构性质。了解样本抗原蛋白的结构性质有助于选择最合适的抗体,待测样本蛋白的结构域和样本在提取和处理过程中是否会变性,蛋白空间构象的改变,会影响抗体的免疫亲和反应。
(5)单多克隆抗体的选择。一般单克隆抗体特异性强,但亲和力相对小,检测抗原灵敏度相对就低;而多克隆抗体特异性稍弱,但抗体的亲和力强,灵敏度高,但易出现非特异性染色(可以通过封闭等避免)。
2. 二抗选择
(1)种属来源。主要根据一抗种属来源而决定二抗来源,如一抗是小鼠来源,那二抗就买抗小鼠的即可(羊、兔等均可)。
(2)标记物的选择。有HRP、Biotin、荧光素等标记物。免疫组化,WB二抗主要选择HRP,Biotin标记的二抗,而免疫荧光染色可按实验需要选择不同荧光素标记的二抗,如FITC、Cy3、PE等。
不过也有用 HRP标记的protein a或g的,前提是你跑的样里没有IgG之类的东西。

