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首页> 外文期刊>The protein journal >Polyclonal Antibody Development Against Purified CC-NBS-LRR like Protein Fragment from Mature Lageneria siceraria Seeds and Immunolocalization
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Polyclonal Antibody Development Against Purified CC-NBS-LRR like Protein Fragment from Mature Lageneria siceraria Seeds and Immunolocalization

机译：纯化成熟番茄种子的CC-NBS-LRR样蛋白片段的多克隆抗体开发及免疫定位

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摘要

CC-NBS-LRR (CNL) plant proteins are related with highly conserved family of disease resistance protein distinguished by a coiled-coil domain, which plays an important role in innate immunity. The present study reports the purification and identification of CNL like protein fragment (CNL-LPF) by two step chromatography and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF/MS), respectively. Furthermore, current study also illustrated the development of polyclonal antibody against purified CNL-LPF, which was used for immunolocalization of CNL-LPF in cytoplasm of cotyledon, using Fluorescence microscopy and Transmission electron microscopy. Lastly, present study also demonstrates in vitro oligomerization of purified CNL-LPF with multiple bands on 4-10 % gradient native-PAGE; each band representing a small fraction of each oligomer population as evident by immunoblots. In conclusion, the current study deals with the purification and polyclonal antibody development against CNL-LPF.
机译：CC-NBS-LRR（CNL）植物蛋白与高度保守的抗病蛋白家族相关，其特征在于卷曲螺旋结构域，其在先天免疫中起重要作用。本研究报告了通过两步色谱法和基质辅助激光解吸电离飞行时间（MALDI-TOF / MS）分别纯化和鉴定CNL样蛋白片段（CNL-LPF）。此外，当前的研究还说明了针对纯化的CNL-LPF的多克隆抗体的开发，该抗体用于荧光子显微镜和透射电子显微镜对子叶细胞质中CNL-LPF的免疫定位。最后，本研究还证明了纯化的CNL-LPF在4-10％的梯度native-PAGE上具有多个条带的体外低聚;如免疫印迹所证实的，每个条带代表每个寡聚物群体的一小部分。总之，当前的研究涉及针对CNL-LPF的纯化和多克隆抗体开发。

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