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首页> 外文期刊>Hybridoma >Titin A-band-specific monoclonal antibody Tit1 5H1.1. Cellular Titin as a centriolar protein in non-muscle cells.
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Titin A-band-specific monoclonal antibody Tit1 5H1.1. Cellular Titin as a centriolar protein in non-muscle cells.

机译：Titin A波段特异性单克隆抗体Tit1 5H1.1。细胞性Titin在非肌肉细胞中作为一种中心粒蛋白。

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摘要

We report the development of a new mouse anti-titin monoclonal antibody, named MAb Tit1 5H1.1, using the synthetic peptide corresponding to an amino acid sequence in the A-band of the titin molecule as immunogen. In the human skeletal muscle, MAb Tit1 5H1.1 reveals a clearly striated staining pattern, reacting with the A-band of the sarcomere. Electrophoretic, immunoblotting, and amino acid sequence analyses with ESI-MS/MS of human skeletal muscle tissue proved the target antigen of MAb Tit1 5H1.1 to be titin. The antibody reacts with titin also in non-muscle cells, producing a punctate pattern in cytoplasm and the nucleus. The most striking finding was a clear reaction of MAb Tit1 5H1.1 with centrioles in all cell types investigated so far. Immunocytochemical co-localization study with ninein-specific antibodies confirmed that the target antigen of MAb Tit1 5H1.1 is a centriole-associated protein. Experiments of the inhibition of synthesis of titin using titin siRNA duplex for the destruction of titin mRNA have shown a decreased staining of centrioles by MAb Tit1 5H1.1 in non-muscle cells and support the proposal that the target antigen of MAb is indeed titin. We suggest this anti-titin monoclonal antibody could be a valuable tool in the study of titin function and its subcellular location, both in muscle and non-muscle cells.
机译：我们报道了一种新的小鼠抗titin单克隆抗体，称为MAb Tit1 5H1.1的开发，使用了与titin分子A带中的氨基酸序列相对应的合成肽作为免疫原。在人的骨骼肌中，MAb Tit1 5H1.1显示出明显的条纹状染色图案，与肌节的A带反应。用ESI-MS / MS对人体骨骼肌组织进行电泳，免疫印迹和氨基酸序列分析，证明MAb Tit1 5H1.1的靶抗原为titin。抗体在非肌肉细胞中也与肌动蛋白反应，在细胞质和细胞核中形成点状模式。迄今为止，最引人注目的发现是在所有研究的细胞类型中，MAb Tit1 5H1.1与中心粒的明确反应。免疫蛋白化学共定位研究与9inin特异性抗体证实，MAb Tit1 5H1.1的靶抗原是一种与中心蛋白相关的蛋白。使用titin siRNA双链体破坏titin mRNA抑制titin合成的实验表明，MAb Tit1 5H1.1在非肌肉细胞中对中心粒的染色降低，并支持MAb的靶抗原确实是titin的提议。我们建议这种抗titin单克隆抗体可能是研究肌肉和非肌肉细胞中titin功能及其亚细胞位置的有价值的工具。

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