How to use ProLong Gold Antifade Reagent with DAPI?

Directions for Use: Following immunostaining, cover samples with ProLong® Gold Antifade Reagent with DAPI #8961 and coverslip. To prevent fading, allow slides to cure in the dark, at room tem- perature for 24 hours, then seal the coverslips with clear lacquer, such as nail polish. Proceed with imaging.

How do you ProLong gold Antifade reagent?

It comes ready-to-use (room temperatures storage and no mixing required)—just apply a drop to the sample, add a coverslip, cure, and image. ProLong Gold Antifade Mountant is not recommended for mounting samples containing fluorescent proteins, such as GFP.

How long does ProLong gold last?

SlowFade® Gold reagent is intended for short-term use (3–4 weeks) only; samples mounted using SlowFade® Gold reagent may degrade over time.

How do you mount with DAPI?

Place slides on a flat surface. – Turn the vial upside down and open the dropper to remove any air bubbles. – Apply 3-4 drops of mounting medium directly on top of the specimen and spread out evenly by tilting slide back and forth or spread evenly with a 0.2 ml plastic pipette tip making sure the tissue is not touched.

How do you make a mounting solution?

Mounting medium can be made with 9 parts of glycerol and 1 part PBS. The pH should be adjusted to between 8.5 and 9.0. This pH has been found to be optimal by many investigators in preventing fluorescein and rhodamine quenching. pH’s above and below this range will lose fluorescence much more quickly.

What is vectashield?

VECTASHIELD Antifade Mounting Medium with DAPI is a unique, stable formula for preserving fluorescence. This medium inhibits rapid photobleaching of fluorescent proteins and fluorescent dyes. VECTASHIELD Mounting Medium with DAPI does not solidify, but remains a liquid on the slide and can be stored without sealing.

How do you make a medium mount?

Mounting medium can be made with 9 parts of glycerol and 1 part PBS. The pH should be adjusted to between 8.5 and 9.0. This pH has been found to be optimal by many investigators in preventing fluorescein and rhodamine quenching.

How much does mounting medium cost?

Use ONLY about 6-8ul of mounting media per 18mm coverslip. The solution should slowly spread to the edges after you place the coverslip onto the media. If you add too much it will leak out the sides and prevent the nail polish from sealing the coverslip to the slide.

How long does DAPI last?

six months
For long-term storage the stock solution can be aliquoted and stored at =–20°C. For short-term storage the solution can be kept at 2–6°C, protected from light. When handled properly, DAPI solutions are stable for at least six months.

How long does mounting media take to dry?

Place an appropriate sized coverslip over sections and let mounting media dry. Slides can be viewed immediately; however, it may take 24 hours for the mounting media to fully harden.

Why is glycerol used for mounting?

Glycerol raises the refractive index of the mounting media, so you’ll get brighter and higher resolution images. The higher the glycerol concentration, the better the fluorescence image but the worse the DIC (Nomarski) image.

Does vectashield expire?

NOTE: This product has a six month expiration date.

When to use prolong gold antifade reagent with DAPI?

ProLong ® Gold Antifade Reagent with DAPI offers enhanced resistance to photobleaching, is premixed and ready to use, and causes little or no quenching of the fluorescent signal. The reagent cures within 24 hours and samples can be saved for months after mounting.

How is actin stained in prolong gold antifade reagent?

Actin is stained with phalloidin-Alexa 594 (red) and nuclei are stained with DAPI (blue). Sample is mounted in ProLong Gold antifade reagent. Storage at room temperature is recommended but can also be stored frozen (-5 to -30°C). Protect from light. Convenient, on-site access to the products you need. Learn more.

How does prolong gold antifade mountant work?

ProLong® Gold Antifade Mountant is a curing mountant that forms a near-perfect optical path (1.47 RI) and allows longer-term storage of the sample. It comes ready-to-use (room temperatures storage and no mixing required)—just apply a drop to the sample, add a coverslip, cure, and image.

Which is prolong live antifade reagent do you use?

Only ProLong Live Antifade Reagent (Cat Nos. P36974 and P36975) is intended for use with live cells. Can I use ProLong Gold, ProLong Diamond, or ProLong Glass mounting medium in a multi-well plate?