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Electron Microscopy Sciences

Technical Data Sheets

EMS Immuno Mount™ DAPI and DABCO™ (IFMDD) Mounting Medium

EMS Catalog #17989-97, 17989-98

Description

Immuno Fluoro mount with antifading agent 1, 4-Diazobicyclo-(2,2,2-octane (DABCO™) is an aqueous non-permanent, low viscosity mounting medium for immunofluorescence. This unique formula prevents rapid photobleaching of FITC, Texas Red, AMCA, Cy2, Cy3, Cy5, Alexa fluoro 488, Alexa fluoro 594, Green fluorescent protein (GFP), tetramethyly rhodamine and Redox. The fluorescence is retained during storage at 4°C in the dark. This medium does not contain phenylenediamine, which destroys immunofluorescence of Cy dyes. This mounting medium is fortified with DAPI which is a counter-stain for DNA. This product is to be used in situ hybridization techniques or other methods where fluorescence of DNA staining is required. DAPI excites at 360nm and emits at 460nm, producing a blue fluorescence. RNA is also stained with DAPI.

Not recommended for phycoethyrin (PE), phycocyanin (PC) and allophycocyanin (APC).

Intended Use

Immunofluorescence, confocal microscopy

Reagent

Ready to use mounting medium for Immunofluorescence

Refractive Index

1.400 ± 0.002 (These numbers apply to these mounting mediums in solution. Refractive indexes change when the water solvent evaporates and mounting media dries on slides. We do not have the means to measure the refractive indexes of dry mounting mediums; however, we expect the numbers to go higher when dried. The refractive index of water is 1.3330.)

Storage

2-8°C, DO NOT FREEZE

Procedure

  1. Bring the vial to room temperature.
  2. Rinse slide to be mounted with distilled or deionized water; touch the edges of slide on a paper towel to remove excess water. Place slides on a flat surface.
  3. Turn the vial upside down and open the dropper to remove any air bubbles. Apply 2-3 drops of mounting medium and apply coverslip carefully without getting air bubbles.
  4. The specimen is ready for visualization under a microscope.
  5. The coverslip may move, so it may be necessary to seal the edges of cover slip with nail polish or any organic medium.
  6. Method for applying Coverslip: Put 1-2 drops of Mounting medium on the specimen. After 3-4 minutes apply coverslip carefully avoiding air bubbles. Put Kimwipes on the top of coverslip, press gently to remove excess mounting medium. With 200 micropipette add organic mounting medium to seal the edges. Incubate at 37°Cfor one hour in the dark to dry organic mounting medium.
  7. Store slide at 2-8°C in the dark.
  8. Removal of Coverslip: Coverslip can be removed before sealing the edges. Soak slide in warm (37°C) distilled or deiononized water for several minutes. Carefully and slowly move the coverslip. Soak in water for an additional few minutes to remove coverslip. Rinse slide several times with warm water to remove all mounting medium. The slide can be remounted again.

Product Information

EMS Immuno Mount™ DAPI and DABCO™ (IFMDD) Mounting Medium