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LacZ/Immunohistochemistry

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Caty784

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Hi!
Please, can anyone explain the mechanism whereby the reporter gene, lacZ, is used in staining a piece of tissue for subsequent visualization of the product of the gene it is connected to? (I know that in the blue/white screening, it cleaves beta-gal and produces blue color, but how about in tissue itself???)
Thank you so much.
Cat

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 Posted Oct 03, 2008, 0:55 AM
samm

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said:

You've already got it! beta-Galactosidase catalyzes the hydrolysis of X-Gal producing a blue precipitate that can be easily visualized under a microscope.
It acts as a reporter to determine the percentage of transfected cells (or a transgenic mouse with appropriate promoters) expressing the lacZ gene. The assay can be completed in 30 minutes, and the blue precipitate can take from 30 minutes to overnight to appear.

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Posted Oct 03, 2008, 15:34 PM
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