C. robusta (formerly known as C. intestinalis type A) adults were purchased from M-Rep. Gamete isolation, fertilization, dechorionation, and embryo incubation were performed as previously published (Christiaen et al., 2009a (link); Christiaen et al., 2009b (link)). The amount of DNA electroporated varied from 10 µg to 90 µg. Animals were reared at 22–24°C. Embryos used for direct visualization of fluorescent markers were fixed in 4% MEM-FA for 30 min, cleared with an PBST-NH4Cl solution (50 mM NH4Cl, 0.15% Triton-X100, 0.05% Tween-20 in 1× PBS), mounted in 50% glycerol supplemented with 2% Dabco 33-LV antifade reagent (Sigma-Aldrich, #290734) and imaged using a Leica SP8 X Confocal microscope.
Dabco 33 lv antifade reagent
Manufactured by Merck Group
Dabco 33-LV is an antifade reagent used in fluorescence microscopy. It helps to prevent photobleaching of fluorescent dyes and proteins, thereby preserving the fluorescence signal during microscopic observation and imaging.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using dabco 33 lv antifade reagent
1
Electroporating Tunicate Embryos for Imaging
2
Ciona robusta Embryo Electroporation
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Ciona robusta (formerly known as Ciona intestinalis type A) adults were purchased from M-Rep, San Diego, CA. Gamete isolation, fertilization, dechorionation, and embryo incubation were performed as previously published 36, 37 The amount of DNA electroporated varied from 10 µg to 90 µg. Animals were reared at 22-24°C. Embryos used for direct visualization of fluorescent markers were fixed in 4% MEM-FA for 30 minutes, cleared with an PBST-NH 4 Cl solution (50mM NH 4 Cl, 0.15% Triton-X100, 0.05% Tween-20 in 1x PBS), mounted in 50% glycerol supplemented with 2% Dabco 33-LV antifade reagent (Sigma-Aldrich, #290734) and imaged using a Leica SP8 X Confocal microscope.
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