Citrate Antigen Retrieval Solution (50X)

柠檬酸钠抗原修复液(50×)具有修复多聚甲醛等固定液引起的蛋白交联，无论石蜡切片、冰冻切片或者细胞爬片，只要是用多聚甲醛、甲醛或其它醛类试剂固定的样品，进行抗原修复都会有效去除蛋白之间的交联，充分暴露抗原表位，从而大大改善免疫染色效果。 操作流程： 1、试剂准备使用前须用蒸馏水或超纯水稀释柠檬酸钠抗原修复液(50×)，配成抗原修复液(1×)。 2、抗原修复 2.1 对于石蜡切片： 2.1.1 脱蜡：切片在二甲苯中脱蜡2×5min。无水乙醇2×5min。90%乙醇2×5min，70％乙醇5min。蒸馏水2×5min。 2.1.2 抗原修复：将切片浸泡于抗原修复液(1×)中，95-100°C加热约10~30min(最佳的加热时间需根据不同的样品和目的蛋白自行摸索)。 注：抗原修复液(1×)使用前需预热到95-100°C。加热可以使用水浴锅或微波炉。如果使用微波炉加热，需注意避免修复液暴沸。随后大约在20~30min冷却至室温。用免疫染色洗涤液洗涤1~2次，每次3~5min。即可进行封闭等后续的免疫染色步骤。 2.2 冰冻切片：用免疫染色洗涤液洗涤切片5min。抗原修复的过程如石蜡切片的抗原修复过程。 2.3 其它样品的抗原修复，可以参考石蜡切片的步骤进行。 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。 Sodium citrate antigen repair solution (50×) has the ability to repair the protein crosslinking caused by fixed liquid such as paraformaldehyde. Whether it is paraffin, frozen section or cell crawl, as long as the sample is fixed with paraformaldehyde, formaldehyde or other aldehyde reagents, the antigen repair will effectively remove the cross-linking between proteins, fully expose the antigen epitope, and greatly improve the immunostaining effect. Procedure: 1. Before the reagent is ready to use, the sodium citrate antigen repair solution (50×) must be diluted with distilled water or ultra-pure water, and the antigen repair solution (1×) must be prepared. 2. Antigen Repair 2.1 For paraffin sections: 2.1.1 Dewaxing: Sections are dewaxed in xylene for 2×5min. Anhydrous ethanol 2×5min. 90% ethanol 2×5min, 70% ethanol 5min. Distilled water 2×5min. 2.1.2 Antigen repair: The section is immersed in antigen repair solution (1×) and heated at 95-100°C for about 10~30min(the best heating time should be determined according to different samples and target proteins). Note: Antigen repair solution (1×) should be preheated to 95-100°C before use. Heat can be done in a water bath or microwave. If using microwave oven heating, pay attention to avoid the repair solution boiling. Then cool to room temperature in about 20 to 30 minutes. Wash with immunostaining detergent 1~2 times, 3~5min each time. Subsequent immunostaining steps such as blocking can be performed. 2.2 Frozen section: The section was washed with immunostaining detergent for 5min. The process of antigen repair such as the process of antigen repair in paraffin sections. 2.3 The antigen repair of other samples can be carried out by referring to the steps of paraffin section. Tips: For your own safety, please take good protection before using the reagent, such as wearing a lab coat, wearing gloves, etc.

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