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In-gel Fluorescence

Visible Protien

Azure Imager c400-600

In-gel fluorescence lets you visualize proteins in the gel without Western blotting

In Gel-Fluorescence has become popular for validating fusion protein generation, protein-protein interaction analysis, and electrophoretic mobility shift assays. Fluorescent fusion proteins can easily be visualized without having to transfer to a membrane for traditional Western analysis (see image below). The fluorescence can be imaged with a digital imager like the c400 or c600.

The c400 is also compatible with Stain-Free™ gels, which incorporate a compound in the gel matrix that binds tryptophan residues when exposed to UV light, generating fluorescent protein bands. These gels save additional time for visualizing protein products, since there is no staining step required.

The AzureRed protein stain can also provide fluorescent staining of total protein in your gel post-electrophoresis, for relative quantitative comparisons. By including known concentration standards and imaging the stained gel, you can get quantitative information about your gel bands.

Related literature:
Imaging In-Gel Fluorescence and Stain-Free™ Gels with the Azure c600 (PDF)
Stain-Free™ Gels as Part of the Western Blotting Workflow: Imaging and Total Protein Normalization (PDF)




Imaging Fluorescent proteins in-gel for a gel-shift assay

Imaging Fluorescent proteins in-gel for a gel-shift assay

The image on the left shows the interaction of Fluorescein-labeled Ubiquitin (FL-Ub) with its binding partners, E1 and E2 Ubiquitin-activating enzymes, over a treatment time course. The upper bands represent ubiquitin in complex with E1 or E2. Fluorescence was detected with the Azure c600 using excitation at 470 nm and emission filter at 497 nm.