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目的:制备兔抗人LYRM1蛋白的多克隆抗体并对其进行初步鉴定。方法:预测LYRM1蛋白的抗原表位,设计并合成出一段由17个氨基酸组成的多肽,将合成肽与载体蛋白钥孔戚血蓝素(KLH)偶联作为抗原,免疫新西兰大白兔,制备抗LYRM1蛋白的多克隆抗体,利用ELISA、Westernblot鉴定其效价及特异性,并将该抗体用于人脂肪组织LYRM1的表达检测。结果:制备了兔抗人LYRM1蛋白的多克隆抗体并证实此抗体效价较高、特异性较强。结论:采用人工合成多肽作为半抗原成功制备了人类肥胖相关新基因LYRM1的多克隆抗体。
Objective: To prepare polyclonal antibody against rabbit LYRM1 protein and preliminary identification. Methods: The epitopes of LYRM1 protein were predicted and a 17-amino acid peptide was designed and synthesized. The synthetic peptide was conjugated with KLH as antigen to immunize New Zealand rabbits to prepare anti-LYRM1 The polyclonal antibody against LYRM1 protein was identified by ELISA and Western blot. The antibody was used to detect the expression of LYRM1 in human adipose tissue. Results: The polyclonal antibody against rabbit LYRM1 protein was prepared and proved to be highly potent and specific. CONCLUSION: Polyclonal antibodies against LYRM1, a novel gene related to human obesity, have been successfully prepared using synthetic peptides as haptens.