Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Using ELISA to Measure GFP Production

(Protocol summary only for purposes of this preview site)

As a reporter, green fluorescent protein (GFP) has primarily been used to visualize the expression of transfected genes and to track proteins fused to GFP. The use of fluorescence intensity as a measure of the abundance of GFP protein is not a sensitive procedure because of the short linear relationship between GFP concentration and its fluorescence intensity. But the availability of specific antibodies with high binding capacity to GFP now permits the development of assays for measuring GFP levels accurately, even at picomolar quantities. For a detailed description of the molecular characteristics of GFP and related fluorescent proteins, see the information panel Fluorescent Proteins.


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