【摘要】 采用硫酸铵盐析、N⁶-（6氨己基）-5′-AMP-Sepharose亲和层析和DEAE-Cellulose离子交换层析,从大鼠睾丸中纯化了LDH-C₄。比活性为16.3 IU/mg,纯化1087倍,产率32.5％。纯化的LDH-C₄经聚丙烯酰胺凝胶电泳（PAGE）鉴定为一条酶带。将纯化的LDH-C₄免疫兔制备了效价为1:64的抗血清。BA-ELISA检测抗血清与大鼠心脏、肌肉和人、小鼠睾丸提取液之间的交叉反应,结果表明抗血清与大鼠心脏、肌肉提取液间存在微弱的交叉反应,而双向免疫扩散未形成沉淀线;抗血清与小鼠睾丸提取液之间存在很强的交叉反应,但与人睾丸提取液之间的交叉反应却很弱,双向免疫扩散显示它们的抗原决定簇部分相同,提示不同种属的LDH-C₄之间抗原性存在差异。更多还原

【Abstract】 Ammoniun gulfate fractionation, N6-(6-aminohexyl)-5’ AMP-Sepharoge affinity ohromatography and DEAE-Celluloge ion-exohange ohromatography were used to extract LDH-C4 from rat testis with a specific activity of 16.3 IU/mg, representing a 32.5% yield and about 1087-fold purification. The purified enzyme appeared to be homogeneous on polyaorylamide gel electrophoresis and a rabbit anti-rat LDH-C4 antigerum was prepared. BA-ELISA was used to examine the corss-reaotivity of the antiaerum with the extract of rat heart and mugole and the extract of human and mice tegtes. Eesultg indicated that there existed a weak cross-reaction between the antigeram and the extract of rat heart and muscle, but no precipitate was found by two dimengional immuno-diffusion. The antibody had a strong crews-reactivity with the extract of mica testig but very weak with that of human testig. Two dimengional immuno-diffusion showed their antigenic determinantg to be partially identical. This indicated that there is some species difference in the determinantg of LDH-C4.更多还原