摘要
为制备兔抗肌球蛋白Ⅹ(myosinⅩ,MyoⅩ)多克隆抗体,合成了含有MyoⅩN端1 000～1 021个氨基酸的多肽,将纯化后的多肽与钥孔戚血蓝蛋白(keyhole limpethemocyanin,KLH)进行交联;通过背部皮下免疫新西兰大白兔获得免疫血清,并通过ELISA、免疫印迹、免疫荧光等分析对免疫血清进行了初步的验证.结果表明:经过4次免疫后可获得免疫血清,ELISA检测表明抗体终效价达到1∶51 200,抗原吸附实验进一步证明了此多克隆抗体的特异性.利用获得的多克隆抗体检测了多种组织中MyoⅩ的表达,发现兔抗MyoⅩ多克隆抗体可特异性识别多种组织中的MyoⅩ.免疫荧光染色表明该抗体可以显示MyoⅩ在COS-7细胞、NLT细胞和神经元中的分布.
To obtain the rabbit polyclonal antibodies against Myosin Ⅹ,a polypeptide containing of 1 000~1 021 amino acids from N-terminal of Myosin Ⅹ was synthesized and coupled to KLH.Then,the polypeptide was used to immunize New Zealand white rabbits by subcutaneous injection to prepare the antiserum.The titer of the antiserum reached to 1∶51 200 after 4 times of immune as revealed by ELISA,and the specificity of the polyclonal antibody was confirmed by antigen absorption test.Furthermore,found this polyclonal antibody can specifically recognized Myo Ⅹ in several tissues as revealed by Western blot.In addition,immunofluorescence displayed the distribution of Myosin Ⅹ in COS-7 cells,NLT cells and primarily cultured neurons.In conclusion,the Myosin Ⅹ polyclonal antibody was successfully prepared,which provided a powerful tool for further study of expression and distribution of Myosin Ⅹ in tissues and cells.
出处
《东北师大学报（自然科学版）》
CAS
CSCD
北大核心
2012年第1期140-144,共5页
Journal of Northeast Normal University(Natural Science Edition)
基金
国家自然科学基金资助项目(30871302
301170142)
吉林省自然科学基金资助项目(200805130)
