LacZ染色试剂盒,LacZ染色

 LacZ染色试剂盒

LacZ Staining Kit  

基本信息：

大肠杆菌中的LacZ基因是应用极广的报告基因之一，被用来检测表达载体介导的基因转移效率，也用于基因启动子研究。LacZ编码β-半乳糖苷酶，该物质非常稳定，能抵抗蛋白酶的水解，并且可以利用许多底物，包括X-gal。X-Gal是β-半乳糖苷酶（β-galactosidase）的显色底物，在β-半乳糖苷酶的催化下会产生蓝色产物，所以可以轻易的检测出LacZ基因的表达。

The LacZ gene from E. coli is one of the most commonly used reporter genes for testing the efficiency of expression vector mediated gene transfer and for studying the regulation of promoters of genes. The LacZ gene encodes the enzyme β-galactosidase, which is very stable, resistant to proteolytic degradation, can utilize a variety of substrates and can be easily assayed in situ. The β-Galactosidase staining kit utilizes X-gal as the substrate.

产品说明：

华越洋的β-半乳糖苷酶染色试剂盒利用X-gal作为底物，配合试剂盒内提供的其他试剂，可以完成250次染色反应。

操作方法:

1.     Remove culture medium and wash cells once with 1 ml of 1X PBS.

2.     Fix the cells with 0.5 ml of Fixative Solution for 10 - 15 min at room temperature.

3.     While the cells in the Fixative Solution, prepare the Staining Solution Mix. Using polypropylene plastic tube only. Prepare enough solution for the number of wells to be stained. For each well, prepare the following mixture:

470 μl of Staining Solution
5   μl of Staining Supplement

25  μl of 20 mg/ml X-gal in DMF

1. Wash the cells two times with 1 ml of 1X PBS.

2. Add 0.5 ml of the Staining Solution Mix to each well. Cover the plate. Incubate overnight at 37℃.

3.  Observe cells under a microscope for development of blue color (200X total magnification).

4. For long-term storage of the stained plates, remove the Staining Solution and overlay the cells with 70 % glycerol. Store at 4℃.

使用注意事项:

• The above protocol is designed for each well in a 12-well culture plate. For using large plate, increase the volume proportionally (e.g., For 6-well plate, double the volume).

• Prepare 1X PBS Solution (not provided). Prepare 3 ml per well.

•  Prepare X-gal Solution: Dissolve 20 mg X-gal in 1 ml DMSO or DMF (N-N- dimethylformamide, not provided) to prepare a 20X stock solution. Excess X- gal solution can be stored at -20℃ (protect from light) for one month. Always use polypropylene container or glass to make and store X-gal. Do not use polystyrene.

• Staining Solution and Staining Supplement: If precipitation occurs, simply warming up the solution to solubilize the precipitates.

保存条件：

• 避光，置于-20℃保存，12个月有效。

  Store kit at 4℃ or -20℃, protected from light.

  Store reconstituted X-gal in -20℃.

  All components supplied are stable for 1 year.