FluoroNanogold

Updated: June 1, 2006 Contents Introduction Features of FluoroNanogold® Applications Alexa Fluor®* 488 FluoroNanogold Alexa Fluor®* 594 FluoroNanogold Fluorescein FluoroNanogold Current Price List Alexa Fluor®* 488 FluoroNanogold Alexa Fluor®* 594 FluoroNanogold Fluorescein FluoroNanogold Introduction 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)).
• Check your labeling by fluorescence before processing for EM (see: Powell, R. D., et al.: J. Histochem. Cytochem., 45, 947-956 (1997)).
• Monitor a dynamic process by fluorescence to determine when to fix and process for EM.
• Differentiate different targets using different fluorophores.

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Alexa Fluor®* 488 FluoroNanogold

• 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-Fab'	Alexa Fluor® 488 FluoroNanogold-Streptavidin
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Product information (PDF)	Product information (PDF)
Material Safety Data Sheet (PDF)	Material Safety Data Sheet (PDF)

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Alexa Fluor®* 594 FluoroNanogold

• 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.

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-Fab'	Alexa Fluor® 594 FluoroNanogold-Streptavidin
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* Alexa Fluor is a registered trademark of Molecular Probes, Inc.

Fluorescein FluoroNanogold

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-Fab'	Fluorescein FluoroNanogold-Streptavidin
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Price List: FluoroNanogold

Alexa Fluor®* 488 FluoroNanogold conjugates

Size	Price
0.5 mL	$241
1.0 mL	$386

7202: FluoroNanogold-anti-mouse Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against mouse IgG (whole molecule).
Fluorophore: Alexa Fluor®* 488.

7204: FluoroNanogold-anti-rabbit Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against rabbit IgG (whole molecule).
Fluorophore: Alexa Fluor®* 488.

7206: FluoroNanogold-anti-goat Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in rabbit, against goat IgG (whole molecule).
Fluorophore: Alexa Fluor®* 488.

7208: FluoroNanogold-anti-rat Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against rat IgG (whole molecule).
Fluorophore: Fluorescein.

7251: FluoroNanogold-anti-sheep Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in rabbit, against sheep IgG (whole molecule).
Fluorophore: Fluorescein.

7253: FluoroNanogold-anti-human Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against human IgG (whole molecule).
Fluorophore: Fluorescein.

7255: FluoroNanogold-anti-guinea pig Fab'-Alexa Fluor®* 488
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against guinea pig IgG (whole molecule).
Fluorophore: Fluorescein.

7216: FluoroNanogold-streptavidin Alexa Fluor®* 488
1.4 nm gold particle is attached to streptavidin.
Fluorophore: Alexa Fluor®* 488.

* Alexa Fluor is a registered trademark of Molecular Probes, Inc.

Alexa Fluor®* 594 FluoroNanogold conjugates

Size	Price
0.5 mL	$241
1.0 mL	$386

7302: FluoroNanogold-anti-mouse Fab'-Alexa Fluor®* 594 (FNG-GAM-Fab'-A594)
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against mouse IgG (whole molecule).
Fluorophore: Alexa Fluor®* 594.

7304: FluoroNanogold-anti-rabbit Fab'-Alexa Fluor®* 594 (FNG-GAR-Fab'-A594)
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against rabbit IgG (whole molecule).
Fluorophore: Alexa Fluor®* 594.

7306: FluoroNanogold-anti-goat Fab'-Alexa Fluor®* 594 (FNG-RAG-Fab'-A594)
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in rabbit, against goat IgG (whole molecule).
Fluorophore: Alexa Fluor®* 594.

7316: FluoroNanogold-streptavidin Alexa Fluor®* 594 (FNG-S-A594)
1.4 nm gold particle is attached to streptavidin.
Fluorophore: Alexa Fluor®* 594.

* Alexa Fluor is a registered trademark of Molecular Probes, Inc.

Fluorescein-FluoroNanogold conjugates

Size	Price
0.5 mL	$241
1.0 mL	$386

7002: FluoroNanogold-anti-mouse Fab'-Fluorescein (FNG-GAM-Fab'-FLSN)
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against mouse IgG (whole molecule).
Fluorophore: Fluorescein.

7004: FluoroNanogold-anti-rabbit Fab'-Fluorescein (FNG-GAR-Fab'-FLSN)
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against rabbit IgG (whole molecule).
Fluorophore: Fluorescein.

7055: FluoroNanogold-anti-guinea pig Fab'-Fluorescein (FNG-GAGP-Fab'-FLSN)
1.4 nm gold particle is attached to affinity-purified Fab' fragment,
raised in goat, against guinea pig IgG (whole molecule).
Fluorophore: Fluorescein.

7016: Fluorescein FluoroNanogold-streptavidin Fluorescein (FNG-S-FLSN)
1.4 nm gold particle is attached to streptavidin.
Fluorophore:Fluorescein.

Custom labeling may be available. Call or Email at tech@nanoprobes.com to inquire.