摘要
为了对精子膜凝集素受体进行定量测定,建立了一种 HRP-凝集素分光光度法,并筛选确定了适宜的测定条件。该方法的主要步骤为:①调节精子浓度;②用 HRP-凝集素孵育精子样品及对照;③加入底物显色;④离心后以分光光度法测定样品和对照上清中游离 HRP-凝集素量。精子膜凝集素受体量以精子样品 OD 值与对照 OD 值之差(即标记到精子膜上的凝集素量)表示。对同一精子样品分别进行三种凝集受体定量的重复试验表明,该法具有良好的再现性。本方法可用于精子膜特性的研究和某些雄性(或男性)不育的诊断。
For quantitative determination of lectin receptors on the sperm surface,an en- zyme-labeled lectin spectrophotometic method was established.The procedure of this method mainly included:①adjusting the density of sperm:②incubating sperm sample with HRP-la- beled lectin to maximal binding:③spectrophotometically determining the unbound lectin in the supernatant after centrifuging the incubated sample.The amount of lectin bound to the spetrm surface was estimated by substracting the OD value of unbound lectin from that of spermless control,which was regarded as the relative amount of lectin-receptors on the sperm surface.The optimal determing conditions of this method were experimentally defined.Both the repeated test and the application of this method to quantitating the lectin-receptors of bull and ram sperm in our research works have been shown that this enzyme-labeled lectin spectrophotometic method is handy and accurate and has a fine repeats.
关键词
精于膜
凝集素受体
酶标凝集素
sperm membrane
lectin-receptor
enzyme-labeled lectin
