Overview:
The activating transcription factor ATF2 (also called CRE-BP1) is a member of the ATF/CREB family of leucine zipper transcription factors, which binds both AP-1 and Cre DNA response element (1). ATF2 has two unique characteristics: it mediates adenovirus E1A-induced trans-activation and forms a heterodimer with c-Jun (2). ATF2 is a target of the JNK signal transduction pathway (3). ATF2 is phosphorylated by JNK on two closely spaced threonine residues located in the NH2-terminal activation domain. The replacement of these phosphorylation sites with alanine inhibited the transcriptional activity of ATF2. Furthermore, p38 MAPK can also phosphorylate ATF2.
References:
1. Maekawa, T. et al: Leucine zipper structure of the protein CRE-BP1 binding to the cyclic AMP response element in brain. EMBO J 1989 8:2023-2028.2. Nomura, N. et al: Isolation and characterization of a novel member of the gene family encoding the cAMP response element binding protein CRE-BP1. J Biol Chem. 1993 Feb 25;268(6):4259-66.3. Gupta, S. et al: Transcription factor ATF2 regulation by the JNK signal transduction pathway. Science, 1995 Jan 20;267(5196):389-93.
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多谢!
不知道能不能直接买到大鼠抗豚鼠血管IgG的ELISA试剂盒。
如果买不到这种试剂盒,用空白板的话(就是反应孔中没有抗原或者抗体包被),把自制的豚鼠血管抗原加到反应孔中进行孵育,能不能使抗原附着在孔壁上?这样的话就可以按照elisa的操作步骤进行大鼠血清检测了。
目前的设想是自制抗原,粉碎豚鼠的血管,制成悬液,通过反复多次的冻融,离心后收集上清液作为抗原(即豚鼠血管抗原)。
然后将抗原加入到elisa的反应孔中(这种是特制的空白板,就是反应孔中没有抗原或抗体包被)进行孵育,使豚鼠血管抗原附着在孔壁上,就是让反应孔充当固相载体,形成固相抗原。倒掉多余的抗原。
再加入待检测的大鼠血清,这样血清中的特异性抗豚鼠血管IgG就可以跟固相的抗原结合,形成固相抗原抗体复合物。
加入酶标的兔抗鼠或者羊抗鼠IgG,形成酶标的抗原抗体复合物。
然后就是一些显色步骤。
现在关键的问题就是抗原加入空白板,能不能形成固相抗原,如果不能跟孔壁附着的话,在后面洗涤的过程中就被洗掉了,那就没办法完成检测了。
请各位大侠给指条明路啊!!!小弟在此多谢了!!!

