Woodley D T, Scheidt V J, Reese M J, Paller A S, Manning T O, Yoshiike T, Briggaman R A
J Invest Dermatol. 1987 Mar;88(3):246-52. doi: 10.1111/1523-1747.ep12465467.
Serum from goats immunized with human type V collagen chains that were cut out of polyacrylamide gels contained an antibody that recognized only type V collagen in an enzyme-linked immunoabsorbent assay and did not label laminin, fibronectin, or types I and IV collagen. Western blot analysis of the antibody showed that its determinant was the alpha 3 (V) chain of type V collagen. Indirect immunofluorescent staining of intact human skin with the antibody produced staining of the dermal blood vessels but not of the dermal-epidermal junction (DEJ). In contrast, both the dermal blood vessels and the DEJ were labeled by the antibody if the skin substrate was first split through the lamina lucida region of the DEJ by incubation in 1 M NaCl solution. Indirect immunoelectron microscopy confirmed the staining pattern found by immunofluorescence and defined the ultrastructural localization of type V collagen in skin. Type V collagen is localized within the DEJ to the lamina lucida region and polar aspects of the basal cell keratinocyte plasma membrane.
从用从聚丙烯酰胺凝胶中切出的人V型胶原链免疫的山羊血清中获得的抗体，在酶联免疫吸附测定中仅识别V型胶原，而不标记层粘连蛋白、纤连蛋白、I型和IV型胶原。对该抗体的蛋白质印迹分析表明，其决定簇是V型胶原的α3（V）链。用该抗体对完整的人皮肤进行间接免疫荧光染色，结果显示真皮血管被染色，而真皮-表皮交界处（DEJ）未被染色。相反，如果首先将皮肤底物在1 M NaCl溶液中孵育，使其通过DEJ的透明层区域分离，则真皮血管和DEJ均被该抗体标记。间接免疫电子显微镜证实了免疫荧光发现的染色模式，并确定了V型胶原在皮肤中的超微结构定位。V型胶原定位于DEJ的透明层区域以及基底细胞角质形成细胞质膜的极性部位。
