title: Preparation and Affinity-Purification of Supervillin Isoform 4 (SV4) Specific Polyclonal Antibodies.
abstract: 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.
authors.affiliation: Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Xueran
authors.initials: X
authors.lastname: Chen
authors.affiliation: Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.affiliation: Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.|Cancer Hospital, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Hongzhi
authors.initials: H
authors.lastname: Wang
authors.affiliation: Cancer Hospital, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Haoran
authors.initials: H
authors.lastname: Yang
authors.affiliation: Cancer Hospital, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Ye
authors.affiliation: Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.|Cancer Hospital, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China.|Department of Urology, The First Affiliated Hospital of Anhui Medical University and Institute of Urology, Anhui Medical University, No. 218, Jixi Road, Hefei, 230022, Anhui, People's Republic of China.
authors.forename: Chaozhao
authors.initials: C
authors.lastname: Liang
authors.affiliation: Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China. z.fang@cmpt.ac.cn.|Cancer Hospital, Hefei Institutes of Physical Science, Chinese Academy of Sciences, No. 350, Shushan Hu Road, Hefei, 230031, Anhui, People's Republic of China. z.fang@cmpt.ac.cn.
authors.forename: Zhiyou
authors.initials: Z
authors.lastname: Fang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000302:isolation & purification* / Q000378:metabolism*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D000906:Antibodies; D008565:Membrane Proteins; D008840:Microfilament Proteins; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; C109413:SVIL protein, human
keywords: E345; Fusion protein; Polyclonal antibody; Supervillin isoform 4