FluoroNanogold™is a new immunoprobe which contains both a fluorescent label and the NanoGold® cluster.
FluoroNanogold™is now available with Alexa Fluor®* 488 or Alexa Fluor®* 594, giving you the benefits of brighter fluorescence, reduced photobleaching, and compatibility with a wider pH range, or in its original formulation with fluorescein as the fluorophore. FluoroNanogold™is offered in 1 mL or the more affordable 0.5 mL size.
By combining gold and fluorescence into one probe, the same specimen may be imaged using both fluorescence microscopy (e.g., with a confocal microscope) and at the ultrastructural level by electron microscopy.
Unlike colloidal golds, which have been found to quench fluorescence, Nanogold® shows little effect. All components are covalently attached to ensure stability and long shelf life. Fab' antibody fragments are much smaller probes than IgG conjugates, and have shown excellent penetration into cells and nuclei. FluoroNanogold™conjugates are chromatographically purified to eliminate any aggregates, free gold or unattached fluorescent molecules.
* Alexa Fluor is a registered trademark of Molecular Probes, Inc.
Features of FluoroNanogold™
Unprecedented correlation between fluorescence and EM data.
Single labeling procedure means less chance for specimen perturbation.
Choice of new fluorophores for brighter fluorescence, lower background and multicolor labeling.
Same excellent penetration as found with NanoGold®: much better than 5 and 10 nm colloidal gold probes.
FluoroNanogold™probes are smaller than IgG conjugates.
Covalent coupling of both labels gives stability and long shelf life.
Conjugates are stable and fluorescent at a wide range of pH and ionic strengths.
Applications
Correlative fluorescence and electron microscopy (see: Robinson, J. M., and Vandré, D.D.: J. Histochem. Cytochem., 45, 631-642 (1997)).
Our new Alexa Fluor® FluoroNanogold™ conjugates will give you greatly enhanced performance compared with conventional fluorophores. Benefit from greatly improved fluorescence properties, combined with a new level of freedom from background and non-specific binding.
Increased fluorescence signal and higher quantum yield - ideal for scarce targets or dynamic systems where exposure needs to be restricted.
Fluorescence remains high and consistent across wider pH range.
Improved solubility means reduced non-specific interactions, lower background and higher signal-to-noise ratios.
Uses fluorescein filter sets.
Available in 1 mL or affordable 0.5 mL sizes.
Left: Structure of Alexa Fluor® 488 and Nanogold® - Fab', showing covalent attachment of components.
Right: Fluorescent staining obtained using combined combined Alexa Fluor® 488 and Nanogold® - Fab' tertiary probe. The specimen is a slide from the NOVA Lite ANA HEp-2 test, an indirect immunofluorescent test system for the screening and semi-quantitative determination of anti-nuclear antibodies (ANA) in human serum (see ). The slide was stained using positive pattern control human sera, a Mouse anti-Human secondary antiboidy, and combined Alexa Fluor® 488 and Nanogold® - Fab' tertiary probe. Specimens were washed with PBS (30 minutes) between each step, then blocked by the addition of 7 % nonfat dried milk to the tertiary antibody solution (original magnification 400 X).
Alexa Fluor® 488 FluoroNanogold™ Conjugates
1.4 nm Nanogold® particle and Alexa Fluor®* 488 fluorophore attached to affinity-purified Fab' fragment.
We can also create custom conjugates, using your own primary antibody (IgG or Fab'), proteins, peptides, lectins, and other molecules. Please use our Custom Synthesis Form to let us know your needs, or drop us a line by phone or email at [email protected].
Our new Alexa Fluor® FluoroNanogold™ conjugates will give you greatly enhanced performance compared with conventional fluorophores. Benefit from greatly improved fluorescence properties, combined with a new level of freedom from background and non-specific binding.
Increased fluorescence signal and higher quantum yield - ideal for scarce targets or dynamic systems where exposure needs to be restricted.
Fluorescence remains high and consistent across wider pH range.
Improved solubility means reduced non-specific interactions, lower background and higher signal-to-noise ratios.
Uses fluorescein filter sets.
Available in 1 mL or affordable 0.5 mL sizes.
Alexa Fluor® 546 FluoroNanogold™ Conjugates
1.4 nm Nanogold® particle and Alexa Fluor®* 546 fluorophore attached to affinity-purified Fab' fragment.
We can also create custom conjugates, using your own primary antibody (IgG or Fab'), proteins, peptides, lectins, and other molecules. Please use our Custom Synthesis Form to let us know your needs, or drop us a line by phone or email at [email protected].
Our Alexa Fluor®* 594 FluoroNanogold™conjugates offer an a second fluorophore, enabling the labeling of more than one target within a specimen with the brightness and photostability of the Alexa Fluor® reagents.
Use for multiple labeling: distinguish a FluoroNanogold™-labeled target from a second target labeled with fluorescein, Alexa Fluor® 488, green fluorescent protein, or other fluorophores.
Uses Texas Red filter sets.
Available in 1 mL or affordable 0.5 mL sizes.
Alexa Fluor® 594 FluoroNanogold™Staining of Caveolin-1a
Localization of caveolin-1a in ultrathin cryosection of human placenta using a new FNG; caveolin 1 alpha is primarily located to caveolae in placental endothelial cells. One-to-one correspondence is found between fluorescent spots and caveola labeled with gold particles (right). Ultrathin cryosections collected on formvar film-coated nickel EM grids were incubated with chicken anti-human caveolin-1a IgY for 30 min at 37°C, then with biotinylated goat anti-chicken F(ab)2 (13 mg/ml) for 30 min at 37°C, then stained with ALEXA-594 FluoroNanogold™-Streptavidin (1:50 dilution) for 30 min at room temperature. Non-specific sites on cryosections were blocked with 1% milk - 5% fetal bovine serum-PBS for 30 minutes at room temperature (figure courtesy of T. Takizawa, Ohio State University, Columbus, OH).
Structure of Alexa Fluor®* 594 FluoroNanogold™Fab' and Streptavidin conjugates, showing covalent attachment.
Alexa Fluor® 594 FluoroNanogold™ Conjugates
1.4 nm Nanogold® particle and Alexa Fluor®* 594 fluorophore attached to affinity-purified Fab' fragment.
We can also create custom conjugates, using your own primary antibody (IgG or Fab'), proteins, peptides, lectins, and other molecules. Please use our Custom Synthesis Form to let us know your needs, or drop us a line by phone or email at [email protected].
Fluorescein is the most widely used fluorophore; fluorescein FluoroNanogold™conjugates combine fluorescein and Nanogold® in a single convenient, stable probe.
correlate fluorescein fluorescence staining with brightfield light microscopy and electron microscopy using a single convenient immunostaining procedure.
Check labeling by fluorescence before EM processing.
Available in 1 mL or affordable 0.5 mL sizes.
FluoroNanogold™Staining of Microtubules
Immunolocalization of microtubules with Fluorescein FluoroNanogold™-Fab' in the same human monocyte visualized by various microscopies: (A) Fluorescence; (B) Phase; (C) DIC, and (D) Electron Microscopy (with silver enhancement). (Micrographs courtesy of Dr. J. M. Robinson and Dr. D. Vandré, Ohio State University).
Fluorescein FluoroNanogold™ Conjugates
1.4 nm Nanogold® particle and Fluorescein fluorophore attached to affinity-purified Fab' fragment.
We can also create custom conjugates, using your own primary antibody (IgG or Fab'), proteins, peptides, lectins, and other molecules. Please use our Custom Synthesis Form to let us know your needs, or drop us a line by phone or email at [email protected].
We can also create custom conjugates, using your own primary antibody (IgG or Fab'), proteins, peptides, lectins, and other molecules. Please use our Custom Synthesis Form to let us know your needs, or drop us a line by phone or email at [email protected].
* Alexa Fluor is a registered trademark of Molecular Probes, Inc.
Alexa Fluor®* 488 FluoroNanogold™![[Alexa Fluor® 488 FluoroNanogold: structure and fluorescence staining] (48k)]](../Images/catfig15.jpg)
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Product information (PDF)
Alexa Fluor®* 546 FluoroNanogold™![[Alexa Fluor® 594 Labeling (64k)]](../Images/catfig17.jpg)
![[Alexa Fluor® 594 Conjugates (46k)]](../Images/catfig16.gif)
![[FluoroNanogold labeling of Microtubules by fluorescent, light and electron microscopy] (302k)](../Images/catfig6.jpg)