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用新鲜人肝癌组织细胞悬液免疫Balb/c小鼠,取小鼠脾细胞与小鼠骨髓瘤细胞系Sp2/0融合,获得两株抗人胶原蛋白单克隆抗体CAb-15和CAb-16。应用Western blotting方法对两株抗体的抗原决定簇进行了鉴定。结果表明,CAb-15的抗原决定簇位于α_1[Ⅲ]链,CAb-16位于Ⅲ型胶原的三螺旋结构区。应用CAb-15和CAb-16对在福尔马林中固定和石蜡包埋的正常肝、肾、主动脉、皮肤、软骨和胎盘等组织进行了免疫组织化学定位。结果表明两株Ⅲ型胶原抗体均可用于免疫组织化学研究。
Balb / c mice were immunized with fresh human hepatoma cell suspension and splenocytes were fused with mouse myeloma cell line Sp2 / 0 to obtain two anti-human collagen monoclonal antibodies CAb-15 and CAb-16. The antigenic determinants of the two antibodies were identified by Western blotting. The results showed that the antigenic determinant of CAb-15 was located in the α_1 [Ⅲ] chain, and CAb-16 was located in the triple helix region of type Ⅲ collagen. Immunohistochemical localization of fixed and paraffin-embedded normal liver, kidney, aorta, skin, cartilage and placenta was performed using CAb-15 and CAb-16 in formalin. The results showed that both type Ⅲ collagen antibodies can be used for immunohistochemistry.