Bennai F, Greney H, Vonthron C, Bousquet P, Dontenwill M
Laboratoire de Pharmacologie Cardiovasculaire et Rénale, CNRS URA 589, Université Louis Pasteur, Faculté de Médecine, Strasbourg, France.
Eur J Pharmacol. 1996 Jun 13;306(1-3):211-8. doi: 10.1016/0014-2999(96)00192-6.
Polyclonal antibodies were raised in rabbits against purified polyclonal anti-idazoxan antibodies. The anti-idiotypic antibodies thus obtained, proved able to inhibit [3H]idazoxan specific binding to anti-idazoxan antibodies. Applied to human nucleus reticularis lateralis membrane preparations, these antibodies (20 micrograms) inhibited about 50 and 70% of the imidazoline specific binding of [3H]idazoxan and [3H]clonidine, respectively. Furthermore, they specifically immunoprecipitated 50% of [3H]idazoxan binding activity of imidazoline binding sites solubilized from the same tissue. [3H]Rauwolscine binding to alpha 2-adrenoceptors in rat cortex was not significantly affected by these antibodies. The antibodies labeled a 43 kDa protein in Western blots of partially purified imidazoline binding sites from human brain. In conclusion, these anti-idiotypic antibodies recognize imidazoline binding sites from human brain and allow the detection of a 43 kDa binding protein associated with or representing the imidazoline receptor expressed in human brain.
用纯化的多克隆抗咪唑克生抗体在兔体内制备多克隆抗体。由此获得的抗独特型抗体能够抑制[3H]咪唑克生与抗咪唑克生抗体的特异性结合。将这些抗体（20微克）应用于人外侧网状核膜制剂时，分别抑制了[3H]咪唑克生和[3H]可乐定咪唑啉特异性结合的约50%和70%。此外，它们特异性免疫沉淀了从同一组织中溶解的咪唑啉结合位点的[3H]咪唑克生结合活性的50%。这些抗体对大鼠皮层中[3H]育亨宾与α2 -肾上腺素能受体的结合没有显著影响。在人脑部分纯化的咪唑啉结合位点的蛋白质印迹中，这些抗体标记了一种43 kDa的蛋白质。总之，这些抗独特型抗体识别来自人脑的咪唑啉结合位点，并能检测到与人脑中表达的咪唑啉受体相关或代表该受体的一种43 kDa结合蛋白。
