Chui S H, Lewis W H, Lam C W, Lai K N
Department of Health Sciences, Hong Kong Polytechnic, Kowloon.
Clin Biochem. 1993 Apr;26(2):97-100. doi: 10.1016/0009-9120(93)90035-5.
A novel method for studying the charge distribution of different subclasses of serum immunoglobulin A (defined by the light chains) is described. Affinity-chromatography purified immunoglobulins were focused in polyacrylamide gel and were then transferred electrophoretically onto nitrocellulose membranes. The transferred immunoglobulins were detected by rabbit antiserum to human kappa (kappa) or lambda (lambda) chain, swine antirabbit IgG, rabbit anti-peroxidase antibodies and peroxidase, together with a substrate solution comprising H2O2 and diaminobenzidine. Finally, the developed membranes were made transparent with Triton-x 114, scanned at 485 nm with a densitometer to obtain quantitation of charge distribution.
本文描述了一种研究血清免疫球蛋白A不同亚类（由轻链定义）电荷分布的新方法。通过亲和色谱纯化的免疫球蛋白在聚丙烯酰胺凝胶中聚焦，然后通过电泳转移到硝酸纤维素膜上。转移的免疫球蛋白通过兔抗人κ（kappa）或λ（lambda）链抗血清、猪抗兔IgG、兔抗过氧化物酶抗体和过氧化物酶，以及包含H2O2和二氨基联苯胺的底物溶液进行检测。最后，用Triton-x 114使显影后的膜透明，用密度计在485nm处扫描以获得电荷分布的定量结果。
