Epifluorescent analysis of HeLa cells using COX IV (3E11) Rabbit mAb (Alexa Fluor® 488 Conjugate) #4853 (green) and ProLong® Gold Antifade Reagent with DAPI (blue pseudocolor).
Confocal immunofluorescent analysis of COS-7 cells, mounted in either glycerol (upper panels) or ProLong® Gold Antifade Reagent #9071 (lower panels), using β-Actin (8H10D10) Mouse mAb #3700 and Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 555 Conjugate) #4409 (red) (0, 3.3, or 10 min of imaging).
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 temperature for 24 hours, then seal the coverslips with clear lacquer, such as nail polish.
Proceed with imaging.
Store at -20°C. ProLong® Gold Antifade Reagent is stable for 12 months when stored protected from light, at or below -20°C.
Storage: ProLong® Gold Antifade Reagent is stable for 6 months when stored, protected from light, at or below -20ºC.
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. This reagent is compatible with most dyes and is appropriate for use in multi-color labeling assays.
The inclusion of DAPI within this mounting reagent eliminates the need for a separate nuclear counterstaining step. 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) is a blue fluorescent DNA dye that targets double-stranded AT clusters in the DNA minor groove (1). One molecule of dye binds to each 3 base pairs of dsDNA and yields an approximate 20-fold fluorescent enhancement (2). The level of DAPI-DNA fluorescence is proportional to DNA content (3).
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.