华越洋⽣生物提供 QQ: 1733351176
LacZ染⾊色试剂盒使⽤用说明书
β-‐Galactosida S taining K it M annual
编号 名称 产品规格
北京华越洋生物RS3601-‐23 LacZ染色试剂盒 250次分析
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-‐galactosida）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 ud reporter genes f or t esting t he e fficiency o f e xpression v ector m ediated g ene t ransfer and for studying the regulation of promoters of genes. The LacZ gene encodes the enzyme β-‐galactosida, which is very stable, resistant to proteolytic d egradation, c an u tilize a v ariety o f s ubstrates a nd c an b e e asily assayed in situ. The β-‐Galactosida 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。
操作⽅方法:
1.Remove c ulture m edium a nd w ash c ells o nce w ith 1 m l o f 1X P BS.
2.Fix the cells with 0.5 ml of Fixative Solution for 10 -‐ 15 min at room
temperature.
3.While t he c ells i n t he F ixative S olution, p repare t he S taining S olution
Mix. U sing p olypropylene p lastic t ube o nly. P repare e nough s olution for the number of wells to be stained. For each well, prepare the
华越洋⽣生物提供 QQ: 1733351176
following m ixture:
470 μl o f S taining S olution
5 μl o f S taining S upplement
25 μl o f 20 m g/ml X-‐gal i n D MF
4.Wash t he c ells t wo t imes w ith 1 m l o f 1X P BS.
5.Add 0.5 ml of the Staining Solution Mix to each well. Cover the plate.
Incubate o vernight a t 37°C.
6. O brve cells under a microscope for development of blue color
(200X t otal m agnification).
7.For long-‐term storage of the stained plates, remove the Staining
Solution a nd o verlay t he c ells w ith 70 % g lycerol. S tore a t 4°C.
使⽤用注意事项:
•The above protocol is designed for each well in a 12-‐well culture plate. F or u sing l arge p late, i ncrea t he v olume p roportionally (e.g., For 6-‐well p late, d ouble t he v olume).
•Prepare 1X P BS S olution (not p rovided). P repare 3 m l p er w ell.
• P repare 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. E xcess X-‐ g al s olution c an b e s tored a t –20°C (protect f rom light) f or o ne m onth. A lways u p olypropylene c ontainer o r g lass t o make a nd s tore X-‐gal. D o n ot u p olystyrene.
•Staining Solution and Staining Supplement: If precipitation occurs, simply w arming u p t he s olution t o s olubilize t he p recipitates.
保存条件：
避光，置于-‐20℃保存，12个月有效。
Store k it a t 4°C o r –20°C, p rotected f rom l ight.
Store r econstituted X-‐gal i n –20°C.
All c omponents s upplied a re s table f or 1 y ear.
华越洋⽣生物提供 QQ: 1733351176
LacZ染⾊色试剂盒，β-半乳糖苷酶染⾊色试剂盒其他相关产品： DNA浓缩试剂盒
DNA凝胶回收试剂盒
SuperLumia H RP底物ECL高敏试剂盒
SuperLumia H RP底物ECL P lus超敏试剂盒
蛋白PAGE胶快染试剂盒
羟甲基化DNA定量试剂盒（荧光法）
羟甲基化DNA定量试剂盒（比色法）
常规组织切片DNA提取试剂盒
血浆/血清DNA提取试剂盒
TuMinute P CR纯化试剂盒
