Smad2/3 Antibody Sampler 试剂盒中的各抗体仅能识别其特异性的靶蛋白,而与其他家族成员没有交叉反应。针对活化态的抗体仅能识别特定位点被磷酸化后的靶蛋白。针对全Smad2, Smad3, 和 Smad4的抗体可以识别其各自内源性的靶蛋白。合成对应人Smad2 Ser465/467和人Smad3 Ser423/425邻近氨基酸残基序列一致的磷酸多肽,免疫动物获得磷酸化特异性单克隆抗体。全Smad2 和 Smad3单克隆抗体通过使用对应小鼠Smad2和Smad3或人Smad2/3的His198邻近氨基酸残基序列的多肽免疫动物获得。使用对应人Smad4 Pro278及其邻近氨基酸残基序列的多肽免疫动物获得多抗。抗体由蛋白A和肽段亲和层析技术纯化得到。Smad2/3 Antibody Sampler试剂盒提供了一种经济的方式检测TGF-β信号通路的靶蛋白。试剂盒含有足以进行4次western blot实验的一抗和二抗。转化生长因子-β (TGF-β)超家族在调控细胞生长,分化和多种生物系统发育过程中发挥了关键作用。总体来说,信号通路是配体诱导丝氨酸/苏氨酸受体激酶寡聚化,同时TGF-β/activin信号通路中和细胞质信号分子Smad2和Smad3磷酸化,骨形成蛋白(BMP)通路是Smad1/5/8磷酸化,激活信号通路。活化的受体磷酸化Smad蛋白的羧基末端会导致它们与通用信号转录子Smad4结合,转位到细胞核内。活化的Smad蛋白通过协同转录因子调控多种生物功能,导致细胞特定状态的转录调控。 Each antibody in the Smad2/3 Antibody Sampler Kit recognizes only its specific target and does not cross-react with other family members. Activation state antibodies detect their intended targets only when phosphorylated at the indicated site. The total Smad2, Smad3, and Smad4 antibodies detect their respective targets at endogenous levels.Phospho-specific monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser465/467 of human Smad2 and Ser423/425 of human Smad3. Total Smad2 and Smad3 monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the amino termini of mouse Smad2 and Smad3 or His198 of human Smad2/3 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the residues surrounding Pro278 of human Smad4. Antibodies are purified by protein A and peptide affinity chromatography.The Smad2/3 Antibody Sampler Kit provides an economical means of detecting target proteins of the TGF-β signaling pathway. The kit contains enough primary and secondary antibodies to perform four western blots with each antibody.Transforming growth factor-β (TGF-β) superfamily signaling plays a critical role in the regulation of cell growth, differentiation, and development in a wide range of biological systems. In general, signaling is initiated with ligand-induced oligomerization of serine/ threonine receptor kinases and phosphorylation of the cytoplasmic signaling molecules Smad2 and Smad3 for the TGF-β/activin pathway, or Smad1/5/8 for the bone morphogenetic protein (BMP) pathway. Carboxy-terminal phosphorylation of Smad proteins by activated receptors results in their partnering with the common signaling transducer Smad4, and translocation to the nucleus. Activated Smad proteins regulate diverse biological effects by partnering with transcription factors resulting in cell-state specific modulation of transcription.