制备抗小鼠Cabs1(Calcium-binding and spermatid-specific protein 1)多克隆抗体,以便进一步利用小鼠模型解析Cabs1蛋白在精子变形与精子功能调控中的功能。以密码子的简并性和偏好性为原则,优化小鼠Cabs1基因编码序列,构建重组表达载...制备抗小鼠Cabs1(Calcium-binding and spermatid-specific protein 1)多克隆抗体,以便进一步利用小鼠模型解析Cabs1蛋白在精子变形与精子功能调控中的功能。以密码子的简并性和偏好性为原则,优化小鼠Cabs1基因编码序列,构建重组表达载体ET-30a(+)/Cabs1,原核表达系统表达重组蛋白,免疫新西兰大白兔获得血清,通过亲和纯化得到Cabs1多克隆抗体。Western Blot、免疫荧光实验验证抗体的应用价值。结果显示,获得了高效价、特异性识别Cabs1的多克隆抗体。Western Blot、免疫荧光实验发现小鼠Cabs1蛋白开始表达于长型精子阶段,在成熟精子中主要定位于顶体和主段。同时进一步发现该抗体还可以识别人Cabs1蛋白。此外,Cabs1抗体也可以识别天然未变性的Cabs1蛋白,用于鉴定和研究Cabs1蛋白的互作复合物。制备的抗体解决了目前商业化的Cabs1抗体均不能识别小鼠来源的Cabs1蛋白的问题。展开更多
Sperm respond to multiple chemical factors derived from the eggs and reorient their migratory direction–a process called chemotaxis.The chemotaxis of sperm in marine invertebrates has been established well.A couple o...Sperm respond to multiple chemical factors derived from the eggs and reorient their migratory direction–a process called chemotaxis.The chemotaxis of sperm in marine invertebrates has been established well.A couple of ion channels and enzymes are involved in the signal transduction pathway of sperm chemotaxis.In contrast,the mechanism of sperm chemotaxis in mammals is still rudimentary.Although much less is known about the chemotaxis of mammalian sperm,the feature of chemotaxis has been utilized to recruit sperm at the optimum functional state,which may improve the efficiency of assisted reproductive technology.In this review,we summarize potent chemoattractants and molecular regulatory mechanism in marine invertebrates and mammals and discuss the application of chemotaxis in clinical treatment.展开更多
文摘制备抗小鼠Cabs1(Calcium-binding and spermatid-specific protein 1)多克隆抗体,以便进一步利用小鼠模型解析Cabs1蛋白在精子变形与精子功能调控中的功能。以密码子的简并性和偏好性为原则,优化小鼠Cabs1基因编码序列,构建重组表达载体ET-30a(+)/Cabs1,原核表达系统表达重组蛋白,免疫新西兰大白兔获得血清,通过亲和纯化得到Cabs1多克隆抗体。Western Blot、免疫荧光实验验证抗体的应用价值。结果显示,获得了高效价、特异性识别Cabs1的多克隆抗体。Western Blot、免疫荧光实验发现小鼠Cabs1蛋白开始表达于长型精子阶段,在成熟精子中主要定位于顶体和主段。同时进一步发现该抗体还可以识别人Cabs1蛋白。此外,Cabs1抗体也可以识别天然未变性的Cabs1蛋白,用于鉴定和研究Cabs1蛋白的互作复合物。制备的抗体解决了目前商业化的Cabs1抗体均不能识别小鼠来源的Cabs1蛋白的问题。
文摘Sperm respond to multiple chemical factors derived from the eggs and reorient their migratory direction–a process called chemotaxis.The chemotaxis of sperm in marine invertebrates has been established well.A couple of ion channels and enzymes are involved in the signal transduction pathway of sperm chemotaxis.In contrast,the mechanism of sperm chemotaxis in mammals is still rudimentary.Although much less is known about the chemotaxis of mammalian sperm,the feature of chemotaxis has been utilized to recruit sperm at the optimum functional state,which may improve the efficiency of assisted reproductive technology.In this review,we summarize potent chemoattractants and molecular regulatory mechanism in marine invertebrates and mammals and discuss the application of chemotaxis in clinical treatment.
