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Post-Translational Modifications

Fluorescence Western Blotting

Azure 400 – 600 Imagers

Fluorescence Western blotting is a great way to detect proteins of a similar molecular weight

Fluorescence is the best option for detecting proteins of extremely similar molecular weight by Western blot. Antibodies that recognize different epitopes can be conjugated to fluorophores with non-overlapping excitation and emission spectra and imaged simultaneously. Imaging on the 400 imager allows for the simultaneous capture of different colors and accurate analysis of both modified and unmodified versions of a protein of interest. After imaging, analysis can be done immediately with the AzureSpot software.

The 500 and 600 offer more fluorescence imaging options.

Related literature:
Three-Color Western Blots with AzureSpectra Antibodies (PDF)
Imaging Three-color Western Blots with the Azure c600 (PDF)

Reagents and Accessories

Reagents and Accessories

For multiplex detection experiments, it can be convenient to use antibodies already validated by your lab; use AzureSpectra fluorescent antibody labeling kit to prepare primary or secondary antibodies conjugated with the fluorophore of your choice.

Azure also provides fluorescent secondary antibodies, fluorescent Western blot buffers, opaque incubation trays, and other products that can help you on your way to getting the best multiplexing data.

Related literature:
Western blotting workflow brochure (PDF)