ProLong Gold Antifade is specifically designed to help protect fluorescent dyes (i.e. Alexa Fluor) from fading but is not recommended with fluorescent proteins (GFP, RFP, mCherry). It comes ready-to-use and stored at room temperature—just apply a drop on the sample, add a coverslip, cure, and image. ProLong Gold Antifade Mountant is a hard-setting liquid mountant applied directly to fluorescently labeled cell or tissue samples on microscope slides. ProLong Gold delivers optimal protection without significantly quenching the initial fluorescence signal to protect fluorescent dyes from fading (photobleaching) during fluorescence microscopy experiments. It is a curing mounting media with a refractive index (RI) of 1.47 that allows longer-term storage of mounted samples. ProLong Gold Antifade Mountant is offered in a ready-to-use 2 mL dropper bottle, or 10 mL bottle and is stored at room temperature. Just apply a drop on the sample, add a coverslip, cure, and image.

Achieve Excellent Refractive Index Matching Without Acrylic Resins
For high-resolution imaging applications, refractive index matching is critical. ProLong Gold Antifade Mountant will cure to an RI of 1.47 and allows you to avoid using traditional acrylic resins (i.e., fingernail polish), which can damage microscope objectives if they make contact. The refractive index determines how much the path of light is bent, or refracted, when entering a material. In microscopy, refractive index differences between specimen and mounting medium should be minimized and can therefore be reduced by using mountants with a higher RI. Optimizing the RI of your experiment will increase the resolution to provide better quality images. Depending on your sample type, thickness, imaging depth and/or staining reagents (e.g. fluorescent proteins or dyes) and whether you need a curing or non-curing mountant, choosing the right mounting media will impact image resolution and should be strongly considered for optimal experimental design.

ProLong Gold Antifade Mountant is not recommended for mounting samples containing fluorescent proteins such as GFP or TagRFP. For superior antifade protection of fluorescent proteins and fluorescent dyes, ProLong Diamond or ProLong Glass Antifade Mountants are recommended. Please note that ProLong Glass mounting media is recommended for samples > 10 microns and all immersion oil imaging applications. For immediate viewing of a sample, choose our non-curing mountant, SlowFade Gold Antifade Mountant.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

DescriptionProLong™ Gold Antifade Mountant, 1 x 10 mL

Green FeaturesLess hazardous, Sustainable packaging

Quantity10 mL

Shipping ConditionRoom Temperature

Product LineProLong™

Product TypeAntifade Mountant

Reagent TypeAntifade Solution

Volume (Metric)10 mL

Unit SizeEach

Contents & Storage

Storage at room temperature is recommended but can also be stored frozen (-5 to -30°C). Protect from light.

Frequently asked questions (FAQs)

What is the difference between ProLong and SlowFade antifade reagents?

Our ProLong antifade reagents dispense as a liquid that will solidify upon the evaporation of water. SlowFade antifade reagents remain liquid. If you are going to image right away and then dispose of your sample, you do not need a mountant that hardens, such as the SlowFade reagents. If you wish to archive your slide for more than a day, you will want a mounting medium that hardens (or “cures”). This hardening will limit the off-rates of various dye-conjugated antibodies and provides a better refractive index. Also, there will be a lower diffusion rate of free radicals, limiting photobleaching.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

If I use ProLong Gold Antifade Mountant to mount my slides, should I seal the edges?

ProLong Gold Antifade Mountant hardens overnight at room temperature. For short-term storage (a couple of weeks) you do not need to seal the edges of the coverslip, and the sample should be stable. Beyond that time, though, some dye conjugates will have an off-rate into the medium, so cold storage is recommended. Sealing the edges will prevent long-term discoloration (golden color) from developing around the edges of the coverslip as the anti-oxidants oxidize.

The edges may be sealed with melted paraffin, VALAP (1:1:1 vaseline, lanolin, paraffin) or epoxy glue. Nail polish is not recommended as various components of nail polish may diffuse into the mountant and quench fluorescent dyes.

Find additional tips, troubleshooting help, and resources within our Cell Imaging Support Center.

Some antifade mounting media stay as liquid whereas others harden. What is the benefit of having one that hardens?

If you are going to image right away and then dispose of your sample, you probably want a mountant that does not harden. If you wish to archive your slide for more than a day, you want a mountant that hardens (or "cures"). This hardening will slow or prevent off-rate of your dye or conjugate and often produces a better refractive index. Secondary sealing is usually not necessary. Also there will be lower diffusion of free radicals, thus limiting photobleaching.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I mounted my cells in ProLong antifade mounting medium, but now I want to go back and re-label them. Is there a way I can unmount the coverslip after it has cured (hardened)?

Yes. Put the slide in a Coplin jar or beaker filled with warm (37oC) PBS buffer and let it sit, no agitation is required. The hardened ProLong mountant will swell and may slide off or be easily dislodged. If cells are adherent to the coverslip, make certain the coverslip side containing the cell or tissue sample does not land face down in the container or become scratched upon handling. Remove the coverslip, wash a couple of times, and proceed with re-staining and re-mounting in new ProLong mountant.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am using ProLong antifade mounting medium. Do I need to let it cure before imaging? Do I need to seal the edges of the coverslip?

You can image before it cures (hardens), and it will still slow photobleaching, but you have to let it cure overnight to get the best refractive index (resolution). There is no need to seal the edges. In fact, if you seal before it cures, it won't cure correctly. If you are archiving the slide for more than a month, though, seal the edges with resin, paraffin or VALAP (1:1:1 vaseline, lanolin, paraffin) after it cures or there may be slight discoloration along the edges over time.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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