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尘香驻风

金虫 (小有名气)

[交流] 萤光素酶检测系统 已有2人参与

有没有虫友用过Promega公司的萤光素酶检测系统试剂盒啊?用的是哪种型号的?
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夏尔list

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我会说我做过么
2楼2014-07-21 18:45:46
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夏尔list

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Dual-luciferase Reporter assay system
3楼2014-07-21 18:48:20
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尘香驻风

金虫 (小有名气)

引用回帖:
3楼: Originally posted by 夏尔list at 2014-07-21 18:48:20
Dual-luciferase Reporter assay system

这是双基因的吧?有没有用过单基因的呀?
4楼2014-07-21 20:42:28
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尘香驻风

金虫 (小有名气)

引用回帖:
3楼: Originally posted by 夏尔list at 2014-07-21 18:48:20
Dual-luciferase Reporter assay system

请问你用这个检测的时候是多样品还是少样品,我看它说明书上说的是发光至少持续一分钟,那么会不会等加完全部的时候前面加的就开始衰退了呢?

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5楼2014-07-22 09:16:43
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引用回帖:
5楼: Originally posted by 尘香驻风 at 2014-07-21 20:16:43
请问你用这个检测的时候是多样品还是少样品,我看它说明书上说的是发光至少持续一分钟,那么会不会等加完全部的时候前面加的就开始衰退了呢?...

These kind of assays are usually done with a Luminescence Microplate Reader. All you need to do is to prepare your cell lysates and add them to a 96-well plate. Then the machine will read the plate for you. The machine reads well by well in the plate and will inject the Stop & Glo reagents right before reading....

You can do it manually if you can't fine the machine.  In this case, you have to add the reagents by yourself and read the samples right away. It's time-consuming though.

If you do such experiments as transient transfection and transactivation assays, you need cotransfect a control reporter (Renilla series). That's the reason that you need a dual-luciferase assay system.
6楼2014-07-22 11:47:10
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