琥珀酸脱氢酶染色液

琥珀酸脱氢酶（succinate dehydrogenase SDH）位于线粒体嵴膜内侧面，与线粒体内膜结合，是三种羧酸循环中**与膜结合的酶，是线粒体的标志酶。SDH酶的活性反应处呈蓝紫色颗粒状，定位分布于心肌细胞和细胞膜，心肌细胞间质不着色。酶活性高时颗粒密集、粗大，青年动物酶活性明显高于幼年、老年动物及心肌细胞缺血、缺氧损伤时，SDH的活性明显降低。 操作流程： 1 样品处理 1.1 对于石蜡切片： 二甲苯中脱蜡 2×5min；无水乙醇 5 min、90%乙醇 2 min、70%乙醇 2 min；PBS 漂洗2 min。 1.2 对于冰冻切片： PBS 漂洗 2 min。 2 琥珀酸脱氢酶染色液染液 2.1 用琥珀酸脱氢酶染色液于湿盒中 37°C，染色 2h； 2.2 流水漂洗切片，2×3~5min； 2.3 用吸水纸吸去切片上的多余水份； 2.4 100%乙醇脱水 3×1~3min； 2.5 二甲苯透明，2×5min，用树脂封片。 染色结果： 心肌 SDH 酶的活性反应处呈蓝紫色颗粒状，分布于心肌细胞内和细胞膜。 注意事项： 1 需一台可观察蓝色荧光的荧光显微镜或激光共聚焦显微镜。 2 需自备 PBS，固定液及抗荧光淬灭封片液。 3 第一次使用本试剂盒时建议先取 1~2 个样品做预实验。 温馨提示： 为了您的自身安全，使用试剂前，请做好防护，如穿实验服，带手套等。 Succinate dehydrogenase (SDH) is located on the inner side of the mitochondrial ridge membrane and binds to the inner membrane of the mitochondria. SDH is the most membrane-binding enzyme among the three carboxylic acid cycles and is the hallmark enzyme of mitochondria. The active reaction site of SDH enzyme was blue-purple granules, located in cardiomyocytes and cell membranes, and the interstitium of cardiomyocytes was not colored. When the enzyme activity was high, the particles were dense and coarse, the enzyme activity of young animals was significantly higher than that of young and old animals, and the activity of SDH was significantly reduced when the myocardial cells were injured by ischemia or hypoxia. Operation process: 1 Sample handling 1.1 For paraffin sections: Dewaxing in xylene for 2×5min; Anhydrous ethanol 5 min, 90% ethanol 2 min, 70% ethanol 2 min; Rinse with PBS for 2 min. 1.2 For frozen sections: Rinse with PBS for 2 min. 2 succinate dehydrogenase stain solution 2.1 Dye succinate dehydrogenase solution in a wet box at 37°C for 2h; 2.2 Rinse slices with running water, 2×3~5min; 2.3 Use absorbent paper to absorb excess water on the section; 2.4 Dehydration with 100% ethanol for 3×1~3min; 2.5xylene transparent, 2 x 5min, sealed with resin. Dyeing result: The active reaction site of myocardial SDH enzyme was blue-purple granule, distributed in myocardial cells and cell membrane. Note: 1 A fluorescence microscope or laser confocal microscope that can observe blue fluorescence is required. 2 Prepare PBS, fixing solution and anti-fluorescence quenching sealing solution. 3 When using this kit for the first time, it is recommended to take 1 to 2 samples for pre-test. Warm reminder: For your own safety, please take good protection before using the reagent, such as wearing a lab coat, gloves, etc.

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