髓鞘发育关键调控因子MYRF多克隆抗体的制备与鉴定
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摘要
MYRF是近年发现的中枢神经系统少突胶质细胞髓鞘化的关键转录因子,但是有效的抗MYRF抗体的缺乏限制了与MYRF相关的髓鞘发育和疾病的研究.通过原核表达和纯化His标签融合的MYRF抗原多肽片段并免疫新西兰兔,制备获得了anti-MYRF多抗.Western Blot、组织和细胞中的免疫荧光等实验进一步确认了该抗体可以特异性地识别小鼠MYRF蛋白.MYRF抗体的成功制备将有助于MYRF相关的髓鞘发育研究.
MYRF is a critical factor of myelin development in the central nervous system (CNS) identified in recent years.However,the lack of efficient anti-MYRF antibodies limits the studies on MYRF-related myelin development and diseases.A method for preparing anti-MYRF polyclonal antibodies by immunizing New Zealand rabbits with bacterial Histagged MYRF peptides is reported.The specificity and efficiency of anti-MYRF antibodies are verified by Western Blot and immunofluorescence in vitro and in vivo.The successful preparation of anti-MYRF antibodies will benefit the researches on MYRF-related myelin development.
MYRF is a critical factor of myelin development in the central nervous system (CNS) identified in recent years.However,the lack of efficient anti-MYRF antibodies limits the studies on MYRF-related myelin development and diseases.A method for preparing anti-MYRF polyclonal antibodies by immunizing New Zealand rabbits with bacterial Histagged MYRF peptides is reported.The specificity and efficiency of anti-MYRF antibodies are verified by Western Blot and immunofluorescence in vitro and in vivo.The successful preparation of anti-MYRF antibodies will benefit the researches on MYRF-related myelin development.
引文
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[9]易敏,杜君卿,黄浩,等.Yap1对少突胶质细胞发育调控的研究[J].杭州师范大学学报(自然科学版),2017,16(3):249-254.
[10]LU Q R,YUK D,ALBERTA J A,et al.Sonic hedgehog-regulated oligodendrocyte lineage genes encoding bHLH proteins in the mammalian central nervous system[J].Neuron,2000,25(2):317-329.
[11]ZHOU Q,WANG S,ANDERSON D J.Identification of a novel family of oligodendrocyte lineage-specific basic helix-loop-helix transcription factors[J].Neuron,2000,25(2):331-343.
[12]HORNIG J,FROB F,VOGL M R,et al.The transcription factors Sox10and Myrf define an essential regulatory network module in differentiating oligodendrocytes[J].PLoS Genetics,2013,9(10):e1003907.
[13]DE STROOPER B,ANNAERT W,CUPERS P,et al.A presenilin-1-dependent gamma-secretase-like protease mediates release of Notch intracellular domain[J].Nature,1999,398(6727):518-522.
[14]BROWN M S,GOLDSTEIN J L.The SREBP pathway:regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor[J].Cell,1997,89(3):331-340.
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