| TechFAQ #3 What precaution should be considered before choosing a mounting medium for Cy2, Cy3, and Cy5 conjugates? |
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| We have been informed recently by Amersham Bioscience, Limited, that aromatic amines,
such as phenylenediamine, can react with cyanine dyes (especially Cy2) and cleave away half of the cyanine molecule. Use of
phenylenediamine as an anti-fading reagent in mounting media may result in weak and diffused fluorescence after storage of the
stained slides. As an alternative, glycerol-based mounting media containing anti-fading agents such as DABCO or n-propyl gallate
may be used. Organic-based mounting media, such as DPX or methyl salicylate, also may be used for cyanine dyes. Methyl salicylate remains liquid so the cover slip needs to be sealed with something like nail polish. DPX will harden into a plastic-like permanent mount. DPX is manufactured by BDH in England and distributed in the U.S. by Gallard-Schlesinger Ind. (800-645-3044). For more information related to the selection and use of secondary antibodies, choose a topic below: |
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