中英文说明书丨细胞衰老β-半乳糖苷酶染色试剂盒

The Cellular Senescence Assay Kit provides an easy-to-use and efficient method to determine cellular

senescence. SA-β-galactosidase catalyzes the hydrolysis of X-gal, which produces a blue color. Each

kit provides sufficient quantities to perform up to 50 assays in 35 mm dishes.

Kit Components

1. 100X Fixing Solution (Part No. 40010): One tube – 1.5 mL of 25% Glutaraldehyde

2. Staining Solution A (Part No. 40011): One tube – 1.5 mL of 500 mM Potassium Ferrocyanide

3. Staining Solution B (Part No. 40012): One tube – 1.5 mL of 500 mM Potassium Ferricyanide

4. Staining Solution C (Part No. 40015): One bottle – 4.5 mL of 1 M Citrate-Na2HPO4 Buffer, pH

6.0, 50 mM MgCl2

5. Staining Solution D (Part No. 40016): One bottle – 4.0 mL of 5 M NaCl

6. X-gal Solution (Part No. 40014): Two tubes – 1.5 mL of 40 mg/mL X-gal in DMF in each tube

Storage

Store X-gal solution protected from light at -20ºC. Store all other components at 4ºC.

Preparation of Reagents

· 1X Fixing Solution: Prepare a 1X Fixing Solution by diluting the provided 100X stock 1:100 in 1X

PBS. Store the diluted solution at room temperature for up to six months.

· Cell Staining Working Solution: Prepare FRESH cell staining working solution based on the

number of samples. The chart below is suggested for samples in 35 mm plate, and may be

modified accordingly to suit other culture plate sizes.

细胞衰老β-半乳糖苷酶染色试剂盒提供了一种有效的方法来可视化衰老相关（SA）ß-半乳糖苷酶。SA-ß-Gal催化X-gal的水解，在衰老细胞中产生蓝色。使用标准光学显微镜可视化结果。

细胞衰老β-半乳糖苷酶染色试剂盒基本参数：

全称：细胞衰老检测试剂盒（细胞衰老相关β-半乳糖苷酶染色法）

英文：Cellular Senescence Detection Kit (SA-β-Gal Staining)

编号：CBA-230

规格：50assays/5 x 50 assays

保存：4ºC

细胞衰老β-半乳糖苷酶染色试剂盒试剂的制备

·1X固定溶液：通过将提供的100X原液稀释成1:100的1X溶液，制备1X固定溶液

PBS。将稀释的溶液在室温下储存六个月。

·细胞染色工作溶液：根据

样本数量。以下图表适用于35mm板中的样品

相应地修改以适合其他培养板尺寸。

分析方案（35mm皿）：

1.从表达SA-ß-Gal的衰老细胞中吸出培养基。

2.用3 mL 1X PBS洗涤细胞两次，并吸出最终洗涤液。

3.加入2mL 1X固定溶液。在室温下孵育5分钟。

4.取出固定溶液，用3mL 1X PBS洗涤固定细胞三次。

5.吸入最后一次洗涤液，并通过添加2 mL新鲜制备的细胞完全覆盖细胞染色工作溶液。

6.在37ºC避光条件下培养细胞4小时至过夜。

7.取出细胞染色工作溶液，然后用3 mL 1X洗涤染色细胞两次PBS并将细胞储存在1X PBS中。对于长期储存，用1X PBS覆盖细胞20%甘油。储存在4ºC。

注：通过将染色样品与DMSO。

8. 使用光学显微镜计数蓝色染色的衰老细胞。

细胞衰老β-半乳糖苷酶染色试剂盒文献参考：

1. Current Protocols in Molecular Biology, John Wiley & Sons Press.

2. Campisi, J. (2000) In Vivo 14, 183-188.

3. Dimri, G. P., X. Lee, G. Basile, M. Acosta, G. Scott, C. Roskelley, E. E. Medrano, M. Linskens, I.

Rubelj, O. Pereira-Smith, M. Peacocke, and J. Campisi. (1995) Proc. Natl. Acad. Sci. USA

92:9363-9367.