CAJ | 학술논문

目的:制备小鼠抗人ASGPR大亚基异构体蛋白H1b的多克隆抗体,并鉴定其特异性.方法:合成H1b特异性多肽,以匙孔血蓝蛋白(KLH)作为载体构建多肽免疫原,通过致敏小鼠,制备小鼠抗人H1b蛋白的多克隆抗体;ELISA法检测抗体效价,通过Western blot和免疫组织化学检测,鉴定抗体的特异性与适用范围.结果:得到小鼠抗人H1b蛋白的多克隆抗体,效价达1:10~5.纯化后的抗体用于Western blot可高特异性识别真核表达的H1b蛋白,并可用于免疫组织化学实验.结论:成功制备小鼠抗人H1b蛋白的多克隆抗体,该抗体具有较高的效价以及特异性,能区分ASG-PR大亚基的两种剪接异构体,从而为进一步研究H1b蛋白的生理功能及其在人类疾病中的意义提供了有利工具.
목적:제비소서항인ASGPR대아기이구체단백H1b적다극륭항체,병감정기특이성.방법:합성H1b특이성다태,이시공혈람단백(KLH)작위재체구건다태면역원,통과치민소서,제비소서항인H1b단백적다극륭항체;ELISA법검측항체효개,통과Western blot화면역조직화학검측,감정항체적특이성여괄용범위.결과:득도소서항인H1b단백적다극륭항체,효개체1:10~5.순화후적항체용우Western blot가고특이성식별진핵표체적H1b단백,병가용우면역조직화학실험.결론:성공제비소서항인H1b단백적다극륭항체,해항체구유교고적효개이급특이성,능구분ASG-PR대아기적량충전접이구체,종이위진일보연구H1b단백적생리공능급기재인류질병중적의의제공료유리공구.
AIM: To prepare and identify mouse polyclonal antibody against protein Hlb, which is the variant of major subunit of human ASGPR. METHODS: Hlb specific peptide was synthesized and coupled with keyhole limpet hemocyanin (KLH) for immunization. Then H1b-KLH conjugation was injected into mouse subcutaneously to produce polyclonal antibody. ELISA assay was used to detect the titer of the antibody. Antibody was also identified by Western blot and immunohistochemistry assays. RESULTS: Mouse antibody against Hlb was prepared after injection of H1bKLH conjugation. The titer of H1b antibody was about 1:10~5.Western blot confirmed its high specificity. This antibody could also be used for immunohistochemistry analysis. CONCLUSION: The successful preparation of the polyclonal antibody against protein H1b, which can discriminate the two variants of the major subunit of ASGPR with high specificity, will provide an efficient reagent for further study of the physiologic functions of H1b and its role in the pathogenesis of human disease.