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作者介绍了一种鉴定蚊虫胃血的直接萤光抗体方法。具体操作如下。抗原:按照Maddy的方法制备人、牛、马、猪及绵羊清除去白细胞的红细胞膜或基质。牛、马的红细胞膜应用丁醇和水溶解分离,将水溶性分馏物干冻并用作抗原;人、猪和绵羊的干冻的整份红细胞膜,用作抗原。上述抗原可在5℃冰筒内长期贮存而不变质。抗血清和结合剂:用白兔进行免疫,使产生抗体血清。初次肌注25毫克干冻基质,或0.5毫升的生理盐水基质匀浆浸出物和0.5毫升的福氏完全佐剂,而后间隔6周再静注25毫克红细胞基质,或0.7毫升盐水基质习浆浸出物2次。每次静注后7天,取兔血约50毫升,分离血清,用40%饱和硫酸铵沉淀2次,取得的球蛋白用异硫氰萤光素或异硫氰四甲基玫瑰红标记。萤光素结合剂为每毫克蛋白质结合约15微克萤光素。玫瑰红结合剂是在球蛋白内加以异硫氰四甲基
The authors describe a method of identifying direct fluorescent antibodies to mosquito stomach blood. The specific operation is as follows. Antigen: Human, cattle, horses, pigs and sheep were prepared according to the method of Maddy to remove leukocyte-depleted erythrocyte membranes or matrices. Erythrocyte membranes of cattle and horses are solubilized and separated using butanol and water, the water-soluble fraction is freeze-dried and used as antigen; the whole red blood cell membrane of human, pig and sheep is used as antigen. The above antigens can be stored at 5 ℃ ice tube long-term without deterioration. Antisera and binding agents: Immunization with white rabbits to produce antibody serum. An initial intramuscular injection of 25 mg of dry matrix or 0.5 ml of saline matrix homogenate extract and 0.5 ml of Freund’s complete adjuvant followed by a further intravenous injection of 25 mg of erythrocyte matrix or 0.7 ml of saline matrix extract 2 Times. Seven days after each intravenous injection, about 50 ml of rabbit blood was obtained. The serum was separated and precipitated twice with 40% saturated ammonium sulfate. The resulting globulin was labeled with isothiocyanate or isothiocyanate. The luciferin incorporates about 15 micrograms of luciferin per milligram of protein. Rose Binding agent is in the globulin to be isothiocyanatotoluene