Heterobifunctional cross-linker인 GMBS를 사용하여 mercaptosuccinylation된 BSA에 putrescine을 결합시켜 토끼에 주사하여 putrescine에 대한 항체를 얻었다. Ouchterlony double immunodiffusion에서 Anti-putrescine serum은 BSA-putrescine과는 침강대를 형성하였으나 BSA과는 침강대를 형성하지 않았다. Anti-putrescine serum은 cadaverine과는 5.3%, spermme과는 3.5% 그리고 spermidine과는 2.3%의 교차 반응성을 보여 주었으나 1,3-diaminopropane과, lysine에 대해서는 교차 반응성이 없었다. Nitrocellulose에 결합시킨 항체와 $[1,4-^{14}C]$putrescine을 이용하여 putrescine에 대한 radioimmunoassay을 개발하였다. 이 측정법에 의한 최저 측정농도는 100 pmol/tube이였다. 대두에서 HPLC에 의한 putrescine치와 TLC에 의한 putrescine치 각각을 RIA에 의한 치에 대하여 상관관계를 조사한 결과 모두 좋은 상관을 보여주었다. 또한 이 측정방법은 putrescine의 유도체로 만들지 않아도 재현성, 특이성 및 정확성이 높았으며 putrescine을 빠르고 간단하게 측정할 수 있었다.
An antibody directed toward putrescine has been produced in rabbits by immunization with a putrescine-protein conjugate which was prepared by a procedure of coupling putrescine to mercaptosuccinylated bovine serum albumin using $N-({gamma}-maleimidobutyryloxy)succinimide$ as a coupling agent. In the analysis of ouchterloney double immunodiffusion, anti-putrescine serum produced precipitation line with BSA-putrescine, but not with BSA. Anti-putrescine serum showed 5.3% crossreaction with cadaverine, 3.5% with spermine, and 2.3% with sper-midine but not crossreactions with 1,3-diaminopropane and lysine. An radioimmunoassay for putrescine has been developed utilizing anti-putrescine serum bound to nitrocellulose and $[1,4-^{14}C]$putrescine. The lower limit of detection by this assay was 100 pmol/tube. In the correlation studies of putrescine determined by this methods (RIA, HPLC, TLC method), RIA showed good correlation with other two methods. This RIA method was turned out to be reproducible, specific, accurate, rapid and simple for determination of putrescine without making derivative of putrescine.
