M Sobue, J Takeuchi, T Fukatsu, T Nagasaka, N Nakashima, T Ogura, T Katoh, K Yoshida
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引用次数: 17
摘要
用软骨素b -裂解酶(0.01单位/ml)在20 mM Tris-HCl (pH 8.0)中处理1小时的组织切片检测硫酸皮肤聚糖蛋白多糖链,然后用n -乙酰半乳糖胺-4硫酸盐偶联不饱和脲酸特异性抗体9A2染色。与此相反,经软骨素b -解酶处理后,抗体3B3和1B5分别与n -乙酰半乳糖胺6-硫酸盐偶联的不饱和脲酸和n -乙酰半乳糖胺偶联的不饱和脲酸反应,未见阳性染色。通过与携带硫酸皮聚糖侧链的小蛋白聚糖反应的单克隆抗体6B6的对比,证实了硫酸皮聚糖的分布。纤维结缔组织阳性染色的定位与这两种方法几乎相同。
Immunohistochemical techniques for detection of dermatan sulfate proteoglycan in tissue sections.
Dermatan sulfate proteoglycan chains were detected in tissue sections treated with chondroitin B-lyase (0.01 units/ml) in 20 mM Tris-HCl (pH 8.0) for 1 hr, followed by staining with antibody 9A2 specific for unsaturated uronic acid coupled to N-acetylgalactosamine-4 sulfate. In contrast, after treatment with chondroitin B-lyase, no positive staining was observed with antibodies 3B3 and 1B5 which react to the unsaturated uronic acid coupled to N-acetylgalactosamine 6-sulfate and unsaturated uronic acid coupled to N-acetylgalactosamine, respectively. The distribution of dermatan sulfate thus revealed was confirmed by comparison with that found by monoclonal antibody 6B6 which reacts with small proteoglycans carrying dermatan sulfate side chains. The localization of positive staining in fibrous connective tissues was almost identical with these two procedures.