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首页> 外文期刊>The protein journal >Preparation and Affinity-Purification of Supervillin Isoform 4 (SV4) Specific Polyclonal Antibodies
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Preparation and Affinity-Purification of Supervillin Isoform 4 (SV4) Specific Polyclonal Antibodies

机译：Supervillin Isoform 4（SV4）特异性多克隆抗体的制备和亲和纯化

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Human Supervillin isoform 4 (SV4), a bigger splicing isoform of Supervillin, contains extra coding exons 3, 4 and 5 (E345), compared to Supervillin isoform 1. Although previous studies have shown that SV4 associated with membrane and cytoskeleton, regulated cell migration and cell survival, its functions are still largely unknown. To broaden our understanding, SV4 specific antibody is important for further study in signaling pathway. The His-SV4 (E345) and GST-SV4 (E345) fusion proteins, which contained SV4 specific domain E345, were purified from bacteria. The His-SV4 (E345) proteins were injected in rabbits as immunogen to produce anti-SV4 serum, and SV4 antibodies were purified by GST-SV4 (E345) proteins cross-linked to affinity resins. SV4 antibodies exclusively recognized SV4 protein both in vitro and in vivo through multi-step testing by ELISA, western blot, immunoprecipitation, and immunofluorescence. Taken together, our data demonstrate a novel SV4-specific polyclonal antibody which will provide a useful tool for further characterization of SV4 function.
机译：人Supervillin同工型4（SV4）是Supervillin的较大剪接同工型，与Supervillin同工型1相比，它包含额外的编码外显子3、4和5（E345）。以及细胞存活，其功能仍然未知。为了拓宽我们的理解，SV4特异性抗体对于信号通路的进一步研究很重要。从细菌中纯化了含有SV4特异性域E345的His-SV4（E345）和GST-SV4（E345）融合蛋白。将His-SV4（E345）蛋白作为免疫原注射到家兔中，以产生抗SV4血清，并通过与亲和树脂交联的GST-SV4（E345）蛋白纯化SV4抗体。 SV4抗体通过ELISA，western印迹，免疫沉淀和免疫荧光的多步测试，在体内外都独家识别SV4蛋白。综上所述，我们的数据表明了一种新颖的SV4特异性多克隆抗体，它将为进一步表征SV4功能提供有用的工具。

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