VECTASHIELD PLUS Antifade Mounting Medium is a unique, stable formula for preserving fluorescence. This medium inhibits rapid photobleaching of fluorescent proteins and fluorescent dyes.
| Unit Size | 2 ml, 10 ml |
|---|---|
| Applications | Immunofluorescence, In situ hybridization |
| Recommended Storage | 2-8 ºC |
| Mounting | Aqueous (Non-Hardening) |
| Antifade | Yes |
| Counterstain | None |
VECTASHIELD PLUS provides several advantages over the regular VECTASHIELD (H-1000 & H-1200). In particular, VECTASHIELD PLUS media have significantly lower inherent background toning and provide superior signal retention particularly in the far-red spectrum (e.g. Cy5).
Similar to other non-setting media, we recommend you to seal the coverslip perimeter if the intention is to store the slides for a period of time.
Yes, the antifade mounting media are at least equivalent in signal retention. We have not seen any reduction in performance with any fluorophores evaluated, and with some dyes (i.e. Alexa Fluor 647 and Dylight 649) there are advantages.
We do know that regular VECTASHIELD antifade mounting media (H-1000 & H-1200) are suitable for SRM. However, the formulation for VECTASHIELD PLUS is different from that used for regular VECTASHIELD. Indeed one component of H-1000 and H-1200 that may have been a contributing factor to the underlying blue tone and that appears to play a key role in optimal signal for SRM, has been removed from VECTASHIELD PLUS. Therefore, until we have data to the contrary VECTASHIELD PLUS would not be recommended for SRM.
The refractive index for VECTASHIELD Mounting Medium is 1.45.
VECTASHIELD PLUS Mounting Medium does not solidify, but rather, remains a liquid on the slide ‐ which can then be stored without sealing. If desired, coverslips can be sealed around the perimeter using nail polish or plastic sealant. Mounted slides should be stored at 2-8 °C and protected from light.
Other manufacturers measure the antifade properties of their mountants using labeled microspheres or arrayed spots. Vector Labs prefers to measure antifade properties of VECTASHIELD mountants using frozen tissue sections that have been immunohistochemically stained with fluorescently labeled secondary antibodies. Antifade capability is then measured using a 40x objective with real time imaging over 30 seconds of continuous exposure to the excitation illumination. Individual intensity measurements are recorded from 6 separate labeled regions and the average is calculated. The intensity after 30 second exposure is expressed as a percentage of the intensity at zero time. The values for PG are taken from the manufacturer's published results.
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