Dylight 647 and 488

Manufactured by Thermo Fisher Scientific

Sourced in United States

DyLight 647 and 488 are fluorescent dyes produced by Thermo Fisher Scientific. DyLight 647 has an excitation maximum at 652 nm and an emission maximum at 672 nm. DyLight 488 has an excitation maximum at 493 nm and an emission maximum at 518 nm. These dyes are commonly used in various biological and biochemical applications that require fluorescent labeling.

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Lab products found in correlation

Prolong diamond antifade reagent, by Thermo Fisher Scientific (3 mentions) Normal goat serum (ngs), by Thermo Fisher Scientific (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Triton x 100, by Merck Group (2 mentions) Millicell ez slide, by Merck Group (2 mentions) Formaldehyde, by Merck Group (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) P iκb α b 9 sc 8404, by Santa Cruz Biotechnology (2 mentions) Cox 2 ma5 14568, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Triton x, by Merck Group (1 mentions) Sz stb1, by Olympus (1 mentions)

Close spelling variants of this product

DyLight 488 (158 citations)

3 protocols using dylight 647 and 488

Zinc Compounds Modulate LPS-Induced Inflammation

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BV-2 cells were seeded at a density of 5 × 10³ cells per well on a Millicell EZ slide (Millipore, Billerica, MA, USA) and grown to 80% confluence in LCM. After 12 h of serum starvation in FBS-free DMEM, cells were treated with Zinc20267861, Zinc18204217, and Zinc33254827 and incubated for 12 h. Following incubation, BV-2 cells were stimulated with 1 μg/ml LPS (Sigma) for 24 h. Samples were fixed with 2% formaldehyde (Sigma-Aldrich) for 10 min, permeabilized with 0.2% Triton-X100 (Sigma) in PBS for 15 min, and then blocked with 5% NGS (Thermo Fisher Scientific) for 1 h at RT. The samples were incubated with COX-2 (MA5-14568; 1:1000; Thermo Fisher Scientific) and p-IκB-α (B-9) (sc-8404; 1:100; Santa Cruz Biotechnology, CA, USA) antibody overnight; then, they were visualized DyLight 647 and 488, 1:1000 (Thermo Fisher Scientific). Sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) 1:5000 (Sigma-Aldrich) and mounted ProLong Diamond antifade reagent (Invitrogen, Thermo Fisher Scientific).

Saddala M.S., Lennikov A., Mukwaya A., Yang Y., Hill M.A., Lagali N, & Huang H. (2020). Discovery of novel L-type voltage-gated calcium channel blockers and application for the prevention of inflammation and angiogenesis. Journal of Neuroinflammation, 17, 132.

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Retinal Pigment Epithelium Immunostaining

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Mice were euthanized by CO2 inhalation. Eyeballs were fixed with 4% paraformaldehyde (Sigma-Aldrich) for 12 h. Under an Olympus SZ-STB1 (Olympus) dissection microscope, the anterior segment tissues, vitreous, and retinas were removed to isolate the retinal pigment epithelia choroid-scleral complex (RCSC). Approximately four to eight relaxing radial incisions were created, and the remaining RCSC were incubated overnight in a blocking solution composed of 5% NGS (Thermo Fisher Scientific) with 0.01% Triton-X (Sigma-Aldrich). The RPE–choroidal-scleral complexes were then incubated with COX-2 (MA5-14568; 1:1000; Thermo Fisher Scientific) and p-IκB-α (B-9) (sc-8404; 1:100; Santa Cruz Biotechnology, CA, USA) antibody for 24 h; following washing three times for 10 min with PBS-T, the samples were incubated for 24 h with DyLight 647 and 488, 1:1000; (Thermo Fisher Scientific) and DAPI 1:5000 (Sigma-Aldrich). Following another wash with PBS-T, the whole mounts were counterstained with DAPI and mounted with ProLong Diamond antifade reagent (Thermo Fisher Scientific).

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Molecular Mechanisms of IMD-0354 in Microglial Inflammation

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BV-2 cells were seeded at 5 × 10³ cells per well in Millicell EZ slides (Millipore, Billerica, MA, USA) and grown to 80% confluence in LCM After 12-hour serum starvation. The cells were treated with 10 and 25 ng/ml IMD-0354; following 1-hour incubation; the cells were stimulated with 1 μg/ml LPS (Sigma, St. Louis, MO, USA) for 24 hours. The samples were fixed with 1% formaldehyde (Sigma, St. Louis, MO, USA) for 10 minutes, permeabilized by incubation with 0.2% Triton X-100 (Sigma, St. Louis, MO, USA) in phosphate-buffered saline (PBS) for 15 minutes, and then blocked with 5% normal goat serum (NGS) (Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at room temperature. The samples were incubated with antibodies against COX-2, p-IκBα (Table 1) overnight; the signals were then visualized using anti-rabbit secondary antibodies DyLight 647 and 488 (1:1000; Thermo Fisher Scientific, Waltham, MA, USA). The sections were counterstained with 4’,6-diamidino-2-phenylindole (DAPI, 1:5000; Sigma, St. Louis, MO) and mounted with ProLong Diamond antifade reagent (Thermo Fisher Scientific, Waltham, MA, USA).

Hikage F., Lennikov A., Mukwaya A., Lachota M., Ida Y., Utheim T.P., Chen D.F., Huang H, & Ohguro H. (2021). NF-κB activation in retinal microglia is involved in the inflammatory and neovascularization signaling in laser-induced choroidal neovascularization in mice. Experimental cell research, 403(1), 112581.

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