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View additional product information for ProLong™ Glass Antifade Mountant - FAQs (P36980, P36984, P36982)
10 product FAQs found
The tissue likely had air trapped in it, either before the labeling process or as a result of air-drying. While the tissue is in blocking solution or another wash, we recommend putting it in a vacuum to pull out internalized air. Air-drying is not necessary; just tap off excess buffer and mount the damp tissue with the mountant.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
ProLong Glass Antifade Mountant (Cat. No. P36980, P36982, P36984) is compatible with any cell or tissue type, ranging in thickness from 0µm to 150 µm, for producing bright, high-resolution Z-stack, 3-D, and 2-D images.
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We offer ProLong Coverslip Sealant (Cat. No. P56128) that can be used to seal the edges of the coverslip and is compatible with both curing and non-curing mountant. The sealant is easy to apply and is brushed on after the mountant has cured, for long preservation of slides. The product page can be found here.
When using a hard-curing mountant, such as ProLong Antifade Mountant, make sure the mountant is fully cured before applying ProLong Coverslip Sealant. Since this sealant can seal moisture under the coverslip, it can interfere with the curing process.
When using a non-curing mountant, such as SlowFade Antifade Mountants, sealing can take place immediately after mounting.
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They are composed of proprietary antifades and polymers in aqueous buffers.
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No. If the sample cannot be sealed with a coverslip, we recommend the use of SlowFade antifade mountant.
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Our ProLong Gold, ProLong Diamond, and ProLong Glass mounting media harden ('cure') by the evaporation of water. Placing the mounted samples in a humid or refrigerated environment would slow down the evaporation process ('curing').
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We have not tested ProLong, ProLong Gold, ProLong Diamond, or ProLong Glass on DyLight dye-conjugated proteins or with other organic dyes, but it is likely that they will be compatible.
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No. ProLong, ProLong Gold, ProLong Diamond, and ProLong Glass reagents are intended for use on fixed or fixed/permeabilized samples. Only ProLong Live Antifade Reagent (Cat Nos. P36974 and P36975) is intended for use with live cells.
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The evaporation of water is the “curing” process for Prolong, ProLong Gold, ProLong Diamond, and ProLong Glass antifade mountant. The evaporation of water causes the polymer to harden. There is no chemical reaction. Please note that there is no curing/hardening for any of the SlowFade reagents or ProLong Live products.
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Bubbles may be removed by one of two methods:
1. Place the amount of ProLong reagent you wish to use on your sample (plus a little excess) in a microcentrifuge tube. Close the cap and centrifuge this aliquot using a tabletop microcentrifuge (speed from 7, 000 to 13,000 rpm). Bubbles should move to the top and these bubbles may be aspirated using a pipettor/pipette tip.
2. Unscrew the lid of the bottle/vial containing the ProLong reagent to make it loose, but do not remove the lid. Place the entire bottle/vial into a vacuum flask, using a faucet aspirator (faucet T-tube). Apply a vacuum (water running through the faucet) and allow vacuum aspiration to occur from 10 to 20 minutes to degas the mixture.
To avoid the formation of bubbles on a sample or to remove bubbles:
1. Before pipetting the desired amount of ProLong reagent for mounting, set the pipettor for a slight excess volume. When pipetting up the mixture, do not pipette up the complete amount, but lift up the pipette tip from the bottle with the pipettor not yet up to full volume. This prevents the aspiration of bubbles into the pipette tip.
2. Bubbles trapped during application of the coverslip: When placing your coverslip onto your drop of ProLong reagent, place the coverslip at a slight angle then, gently lower the coverslip. If the coverslip is lowered flat onto the sample, or lowered too quickly, bubbles can be trapped.
3. Bubbles trapped in tissue: One problem with tissue sections, particularly cryosections is that air can get trapped within and under the section. Upon mounting, bubbles are not observed but as the mountant hardens, it compresses the sample slightly, forcing air out of the section. This leads to microscopic bubbles forming over the section, trapped within the mountant. To avoid this, degas the tissue sample prior to mounting. Place the sections submerged in buffer or blocking solution, into a vacuum chamber and expose the sample to the vacuum. This will degas the sections and buffer. Remove the sample from this degassed buffer and mount.
4. If ProLong-mounted samples have already cured but have bubbles, you can un-mount your sample by placing the slides into PBS (Coplin jar or a Petri dish filled with PBS). The ProLong reagent will swell and the coverslip will slide off or can be gently removed manually. You can then re-mount with a new aliquot of ProLong reagent.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.