T-Pro Biotechnology


for your life science lab

Mounting Reagent

T-Pro Mounting Reagent or T-Pro Mounting Long Reagent is an effective antifade reagent for the immuno-fluorescence staining. This antifade reagent offers enhanced resistance to photobleaching and is premixed and ready to use. The sample mounted by “T-Pro Mounting Reagent or T-Pro Mounting Long Reagent” can be saved for a month after mounting. “T-Pro Mounting Reagent or T-Pro Mounting Long Reagent” offers excellent compatibility with a multitude of dyes and dye complexes
Considerations
• Warm the bottle of “T-Pro Mounting Reagent or T-Pro Mounting Long Reagent” to room temperature before use.
• Remove excess moisture from the slide before “T-Pro Mounting Reagent or T-Pro Mounting Long Reagent” is added.
• T-Pro Mounting Reagent or T-Pro Mounting Long Reagent is ideal for use with most fixed samples in both DIC, phase and fluorescence microscopy.
 
Procedure
1. Warm reagent. Remove the T-Pro Mounting Reagent or T-Pro Mounting Long Reagent from the freezer and allow the vial to equilibrate to room temperature. Do not use an external heat source to warm the T-Pro Mounting Reagent or T-Pro Mounting Long Reagent, as this may decrease the long-term stability of this product.
2. Apply T-Pro Mounting Reagent or T-Pro Mounting Long Reagent. Remove any excess liquid from the specimen by tapping the side of the slide or coverslip on to a clean laboratory wipe and apply 1 drop (10-20 ul) of the “T-Pro Mounting Reagent or T-Pro Mounting Long Reagent” to the specimen. Cover slide-mounted specimens with a coverslip and carefully lower the coverslip onto the antifade reagent to avoid trapping any air bubbles.
3. Prepare slides for viewing. For samples mounted using“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent”, allow the preparation to cure on a flat surface in the “dark”. Curing time may vary from a couple of hours to overnight, depending on the thickness of the sample and the relative humidity of the surrounding air. We recommend 24 hours curing time. For long-term storage, seal the coverslip and store the slide upright in a covered slide box at 4˚C. Desiccant may be added to the box to ensure that the slide remains dry.

 

Figure 1. T-Pro Mounting Long Reagent I provides enhanced resistance to photobleaching. a, Fluorescein-phalloidin were used to stain NIH 3T3 cell and mounted in the present or absent of “T-Pro Mounting Long Reagent I” and illuminated by using a 100-watt Hg-arc lamp for different time as indicated. Samples were imaged using a 63x/1.4 NA oil immersion lens and acquired using a 16-bit monochrome CCD camera. The same exposure settings were used for all images. b, The decreasing rate of fluorescence intensity in different time points were analyze by using metamorph software and shown in panel b.
 

 

Fig. NIH 3T3 cells stained with T-Pro Mounting Long Reagent I (with DAPI), Alexa Fluo 488 phalloidin and Alexa Fluo 594 wheat germ agglutinin.