计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
本试剂盒采用高灵敏度的银染染料，可应用于变性胶及非变性胶的蛋白染色，具 有目的条带清晰、背景低、操作时间可灵活控制的优点。另外，本试剂盒增加了一步 短时敏化作用步骤，可明显降低背景及提升目的条带的亮度。
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注意事项
1．请提前准备50 ml固定液（超纯水：乙醇：冰醋酸=6:3:1）、50 ml洗脱液（10%乙醇） 和50 ml终止液（5%的冰醋酸）。
2．操作时请使用去离子水和洁净的玻璃或塑料器皿，须戴一次性手套进行操作。
3．整个银染过程需在摇床上进行，摇床转速约60 rpm左右。
4．需自备乙醇，冰醋酸。
使用说明
以下操作步骤中各溶液的用量以大小为8.5×5.5 cm、厚度为1.0 mm的凝胶为例，以 凝胶全部浸没到溶液为准，置于摇床上操作，一般用量25 ml。对于大型凝胶，各溶液 的使用量需按凝胶体积的比列放大。 请提前准备好50 ml固定液（超纯水：乙醇：冰醋酸=6:3:1）、50 ml洗脱液（10%乙醇）和50 ml终 止液（5%的冰醋酸）。
1．水洗：电泳完成后，用超纯水洗胶2次，每次洗涤5分钟。
2．固定：用25 ml固定液固定凝胶2次，每次固定15分钟。
3．洗脱：用洗脱液洗胶2次，每次洗涤5分钟。
4．水洗：用超纯水洗胶2次，每次洗涤5分钟。
5．增敏：将上一步洗好的凝胶置于银染增敏工作液中，室温下准确孵育1分钟后用超纯 水洗胶3次，每次洗涤20秒。 银染增敏工作液的配制：取50 µl Silver Stain Sensitizer（500×）加入到25 ml超纯水中，混匀。
6．银染：弃去超纯水，将凝胶置于银染工作液中孵育30分钟。 银染工作液的配制：取25ml Silver Stain加入50 µl Silver Stain Enhancer混匀。
7．水洗：用超纯水快速洗胶2次，每次洗涤准确控制为20秒。
8．显影：立即将洗好的凝胶浸没在显影液中，室温孵育2-3分钟，直至蛋白条带显示清 晰。 显影液的配制：取25ml Silver Stain Developer加入30µl Silver Stain Enhancer混匀。 注意：显影30秒内，蛋白条带开始显现，继续显影至2-3分钟。若蛋白条带显色较浅，可适当延长显 影时间至5分钟及以上。
9．终止：用终止液洗去凝胶上的显影液后，将凝胶浸泡在新的终止液中反应10分钟。
实验图片
BSA蛋白样品经过10%的SDS-PAGE 凝胶电泳后银染结果
BSA蛋白分子量约为66 kD，上样量从
左到右分别为50 ng，10 ng，5 ng
This reagent kit uses highly sensitive silver dye, which can be applied to protein staining of denatured and non denatured gels. It has the advantages of clear target bands, low background, and flexible control of operation time. In addition, this reagent kit has added a short-term sensitization step, which can significantly reduce the background and enhance the brightness of the target band.
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Matters needing attention
1. Please prepare 50 ml of fixed solution (ultrapure water: ethanol: acetic acid=6:3:1), 50 ml of eluent (10% ethanol), and 50 ml of termination solution (5% acetic acid) in advance.
2. Please use deionized water and clean glass or plastic containers during operation, and wear disposable gloves for operation.
The entire silver dyeing process needs to be carried out on a shaker, with a rotation speed of about 60 rpm.
4. Self prepared ethanol and glacial acetic acid are required.
Instructions for use
The dosage of each solution in the following operation steps takes the gel with a size of 8.5 × 5.5 cm and a thickness of 1.0 mm as an example. The gel is immersed in the solution completely, and is operated on a shaker, with a general dosage of 25 ml. For large gel, the dosage of each solution should be scaled up according to the gel volume. Please prepare 50 ml of fixed solution (ultrapure water: ethanol: glacial acetic acid=6:3:1), 50 ml of eluent (10% ethanol), and 50 ml of termination solution (5% glacial acetic acid) in advance.
1. Water washing: After electrophoresis is completed, wash the gel twice with ultrapure water for 5 minutes each time.
2. Fixation: Fix the gel twice with 25 ml of fixative solution for 15 minutes each time.
3. Elution: Wash the adhesive twice with eluent, each time for 5 minutes.
4. Water washing: Wash the glue twice with ultrapure water, each time for 5 minutes.
5. Sensitization: put the gel washed in the previous step into the silver dye sensitization working solution, incubate it accurately for 1 minute at room temperature, and then wash it with ultrapure water for three times, each time for 20 seconds. Preparation of silver staining sensitization working solution: Take 50 µ l Silver Stain Sensitivity (500 x) and add it to 25 ml of ultrapure water, mix well.
6. Silver staining: discard ultrapure water and incubate gel in silver staining working solution for 30 minutes. Preparation of silver staining working solution: Take 25ml Silver Stain and add 50 µ l Silver Stain Enhanced to mix well.
7. Water washing: Quickly wash the glue twice with ultrapure water, with each washing accurately controlled for 20 seconds.
8. Development: Immerse the washed gel in the developer immediately and incubate it at room temperature for 2-3 minutes until the protein strip is clear. Preparation of developer: Take 25ml Silver Stain Developer and add 30 µ l Silver Stain Enhanced to mix well. Attention: Within 30 seconds of development, protein bands begin to appear and continue to develop for 2-3 minutes. If the protein band appears lighter, the development time can be appropriately extended to 5 minutes or more.
9. Termination: After washing the developer on the gel with the termination solution, soak the gel in a new termination solution to react for 10 minutes.
Experimental images
Silver staining results of BSA protein samples after 10% SDS-PAGE gel electrophoresis
The molecular weight of BSA protein is about 66 kD, and the loading amounts from left to right are 50 ng, 10 ng, and 5 ng, respectively
规格
英文名称
Protein Silver Stain Kit
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