Long-Term Photostability of FITC-Channel Fluorescent Conjugated Antibodies

The use of fluorophore labeled antibodies is a powerful tool in biological research. These fluorophore-antibody conjugates have found important applications in Western blotting, ELISA, FACS and fluorescence microscopy.

Because of their prominence in biological assays, a great deal of research has been conducted to identify and synthesize the ideal fluorophore for a given application. While subject to some debate, ideal fluorophores typically possess one or more of the following qualities:

1. Strong fluorescence, as characterized by a high extinction coefficient and quantum yield
2. High photostability and resistance to photobleaching
3. Resilience under different environmental conditions, such as pH
4. Minimal auto-fluorescence or background interference
5. Simplicity and consistency in experimental usage.

Selecting the correct conjugate for your experiment is an early yet crucial part of obtaining satisfactory data. Contact us for any assistance in choosing the optimal probe or labeled antibody from our catalog.

Comparison of Photobleaching Rates: Tubulin of HeLa cells was labeled with (left) conventional fluorescein (FITC) goat anti-mouse IgG or (right) iFluor® 488 goat anti-mouse IgG. Images were taken every 60 seconds on a fluorescence microscope.

Resources & Information

AAT Bioquest has a wide range of resources and specialized products, from Application Notes & FAQs to spectral profiles.

Related Digital Catalogs:
• Antibody and Protein Labeling
• iFluor® Dyes and Kits
• Secondary Antibodies

AssayWise Newsletter Research Articles:
• Brighter Labeling with iFluor® Conjugates
• Quantitative comparison of iFluor® 488- and FITC- conjugated antibodies for use in cell labeling

Multicolor Fluorescent Staining: HeLa cells were stained with mouse anti-tubulin followed by iFluor® 633 Goat Anti-Mouse IgG (red), actin filaments with Phalloidin- iFluor® 488 Conjugate (green), and nuclei with Hoechst 33342 (blue).