
Overview:
NIK is a mitogen-activated protein kinase kinase kinase 14 (MAP3K14), which binds to TRAF2 and stimulates NF-kappaB activity. NIK shares sequence similarity with several other MAPKK kinases and participates in NF-kappaB-inducing signalling cascade common to receptors of the tumour-necrosis/nerve-growth factor (TNF/NGF) family and to the interleukin-1 type-I receptor. (1) NIK is expressed in primary human cells and in inflamed rheumatoid arthritis tissue and plays a selective role in signaling by the lymphotoxin-beta receptor (2). NIK is a therapeutic target in the immune and bone-destructive components of inflammatory arthritis.
References:
1. Smith, C. et.al: NF-kappa-B-inducing kinase is dispensable for activation of NF-kappa-B in inflammatory settings but essential for lymphotoxin beta receptor activation of NF-kappa-B in primary human fibroblasts. J. Immun. 167: 5895-5903, 2001.2. Yin, L. et.al: Defective lymphotoxin-beta receptor-induced NF-kappa-B transcriptional activity in NIK-deficient mice. Science 291: 2162-2165, 2001.
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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!!!
多谢!

