Mack D O, Reed V L, Smith L D
H. L. Snyder Memorial Research Foundation, Winfield, Kansas 67156.
Anal Biochem. 1987 Nov 15;167(1):53-61. doi: 10.1016/0003-2697(87)90133-3.
Agarose gel isoelectrofocusing is used to separate the isozymes of human prostatic acid phosphatase with retention of enzyme activity. The native blotting of the isozymes onto a nitrocellulose membrane increases the sensitivity of the enzyme stain and is suitable for analysis of isozymes in prostate tissue, which contains little nonprostatic acid phosphatase. The specificity of the transfer is increased by treating the membrane with antibody to human prostatic acid phosphatase prior to the transfer. The specificity of the antibody is conferred to the membrane resulting in a transfer specific for prostatic acid phosphatase. The immunotransfer procedure is applicable to serum which contains appreciable amounts of nonprostatic acid phosphatase.
琼脂糖凝胶等电聚焦用于分离人前列腺酸性磷酸酶的同工酶，并保留酶活性。将同工酶原位印迹到硝酸纤维素膜上可提高酶染色的灵敏度，适用于分析前列腺组织中的同工酶，因为该组织中非前列腺酸性磷酸酶含量很少。在转移前用抗人前列腺酸性磷酸酶抗体处理膜，可提高转移的特异性。抗体的特异性赋予了膜，从而实现对前列腺酸性磷酸酶的特异性转移。免疫转移程序适用于含有大量非前列腺酸性磷酸酶的血清。
