Mattson D H, Roos R P, Arnason B G
J Neurosci Methods. 1980 Oct;3(1):67-75. doi: 10.1016/0165-0270(80)90035-7.
An isoelectric focusing (IEF) technique using direct peroxidase or peroxidase-antiperoxidase (PAP) overlay staining for detection of cerebrospinal fluid (CSF) oligoclonal IgG is described. The method involves chemical fixation of all proteins in the gel followed by application of peroxidase coupled antiserum as a specific stain. The direct peroxidase system is 4 times and the PAP system 8 times as sensitive as Coomassie blue staining after standard IEF or Coomassie blue staining of immunofixed gels. As little as 0.5 microgram (direct peroxidase) or 0.3 microgram (PAP) of IgG can be detected. Direct peroxidase staining of IgG in IEF gels can be used to examine unconcentrated CSF, with banding patterns similar to those obtained by Coomassie blue staining after standard IEF of concentrated CSF.
本文描述了一种等电聚焦（IEF）技术，该技术使用直接过氧化物酶或过氧化物酶-抗过氧化物酶（PAP）叠加染色法来检测脑脊液（CSF）中的寡克隆IgG。该方法包括对凝胶中的所有蛋白质进行化学固定，然后应用与过氧化物酶偶联的抗血清作为特异性染色剂。在标准IEF或免疫固定凝胶的考马斯亮蓝染色后，直接过氧化物酶系统的灵敏度是考马斯亮蓝染色的4倍，PAP系统的灵敏度是考马斯亮蓝染色的8倍。最低可检测到0.5微克（直接过氧化物酶）或0.3微克（PAP）的IgG。IEF凝胶中IgG的直接过氧化物酶染色可用于检测未浓缩的脑脊液，其条带模式与浓缩脑脊液标准IEF后考马斯亮蓝染色所获得的条带模式相似。
