Schmidt-Kastner R, Meller D, Bellander B M, Strömberg I, Olson L, Ingvar M
Department of Clinical Neurophysiology, Karolinska Hospital, Stockholm, Sweden.
J Neurosci Methods. 1993 Feb;46(2):121-32. doi: 10.1016/0165-0270(93)90147-j.
Disturbances of the blood-brain barrier (BBB) following brain lesions lead to extravasation of serum proteins that can be detected by immunohistochemical methods in tissue sections. Here, extravasated immunoglobulins were visualized by a 1-step technique using rabbit anti-rat immunoglobulins conjugated to horseradish peroxidase (HRP). This method is associated with a lower background staining than the conventional 3-step peroxidase-antiperoxidase (PAP) technique using rabbit antibodies against rat whole-serum proteins or immunoglobulins (IgG). Further tests using a direct conjugate of rabbit anti-rat immunoglobulins to fluorescein isothiocyanate (FITC) showed usefulness of the approach for fluorescence microscopy. Additional experiments showed that antibodies directed against mouse immunoglobulins as used for detection of mouse monoclonal antibodies can cross-react with extravasated rat immunoglobulins. Therefore, immunohistochemical studies on lesioned rat brain should routinely include a visualization of areas containing extravasated serum proteins including immunoglobulins.
脑损伤后血脑屏障（BBB）的破坏会导致血清蛋白外渗，这可以通过免疫组织化学方法在组织切片中检测到。在此，使用与辣根过氧化物酶（HRP）偶联的兔抗大鼠免疫球蛋白的一步法来观察外渗的免疫球蛋白。与使用兔抗大鼠全血清蛋白或免疫球蛋白（IgG）抗体的传统三步过氧化物酶抗过氧化物酶（PAP）技术相比，该方法的背景染色更低。使用兔抗大鼠免疫球蛋白与异硫氰酸荧光素（FITC）的直接偶联物进行的进一步测试表明，该方法可用于荧光显微镜检查。额外的实验表明，用于检测小鼠单克隆抗体的抗小鼠免疫球蛋白抗体可与外渗的大鼠免疫球蛋白发生交叉反应。因此，对损伤大鼠脑进行免疫组织化学研究时，应常规对包含外渗血清蛋白（包括免疫球蛋白）的区域进行可视化。
