Schalken J J, De Grip W J
Exp Eye Res. 1986 Sep;43(3):431-9. doi: 10.1016/s0014-4835(86)80078-1.
A versatile, multispecies enzyme-linked immunosorbent assay for the rod visual pigment (rhod)opsin has been developed. For this quantitative inhibition assay a monospecific polyclonal antiserum is used which is elicited in rabbits against bovine rod outer-segment membranes. Detergent concentrations as high as 1.0% can be used in the assay with only a slight loss in sensitivity. The assay allows quantitative determination of the apoprotein opsin with a detection level of about 0.04 pmol per sample by using standards prepared by illumination of spectrophotometrically determined amounts of rhodopsin. The antiserum shows considerable cross-reactivity with opsin from several species (mouse, rat, quail, monkey and man). The high degree of monospecificity and cross-reactivity of the antiserum already allowed quantitation of opsin content in crude eye extracts of mouse, rat and quail with a sensitivity comparable to that of bovine opsin. Similar types of multispecies immunoassays for quantitation of highly conserved membrane proteins can be developed using the described approach, requiring only a monospecific antiserum elicited against an easily accessible species and crude tissue extracts both for coating and as a source of the inhibitory antigen.
已开发出一种用于视杆细胞视色素（视紫红质）的通用多物种酶联免疫吸附测定法。对于这种定量抑制测定，使用的是一种单特异性多克隆抗血清，它是在兔子体内针对牛视杆细胞外段膜产生的。该测定中可使用高达1.0%的去污剂浓度，灵敏度仅有轻微损失。通过使用由分光光度法测定的视紫红质光照量制备的标准品，该测定法能够定量测定脱辅基蛋白视蛋白，检测水平约为每个样品0.04皮摩尔。该抗血清与来自几种物种（小鼠、大鼠、鹌鹑、猴子和人类）的视蛋白具有相当大的交叉反应性。该抗血清的高度单特异性和交叉反应性已经能够定量小鼠、大鼠和鹌鹑的粗眼提取物中的视蛋白含量，其灵敏度与牛视蛋白相当。使用所述方法可以开发类似类型的用于定量高度保守膜蛋白的多物种免疫测定法，只需要一种针对易于获取的物种产生的单特异性抗血清以及用于包被和作为抑制性抗原来源的粗组织提取物。
