Co-injected fluorescein dextran (70,000 MW, ThermoFisher, D1822) was used as lineage tracer for dorsal lips in IF analyses. For IF analyses, embryos were fixed in 4% paraformaldehyde for 1 h at RT, followed by two washes in 1× PBS for 15 min each. For staining of animal caps or bisected specimens, embryos were manually dissected horizontally or sagittaly after fixation, transferred to 24-well plates, and washed twice for 15 min in PBST (PBS/0.1% Triton X-100). After blocking for 2 h at RT in CAS-Block (1:10 in PBST; ThermoFisher, #008120) blocking reagent was replaced by antibody solution (diluted in CAS-Block) for incubation ON at 4°C. Antibodies used were: β1-Integrin (DSHB 8C8-s; 1:70), MZ15 (DSHB; 1:200). Then antibody solution was removed and explants washed twice for 15 min in PBS. Secondary antibodies (ThermoFisher, all 1:1000 in CAS-Block) were incubated for 2 h at RT. Cell borders were visualized using AlexaFluorTMPlus 405 Phalloidin (ThermoFisher A30104) overnight at 4°C (1:100 in PBS-). For photo documentation, bisected embryos or caps were transferred onto microscope slides or positioned in low melt agarose on a Petri dish (0.5% low melt agarose in 1× PBS-).
Co injected fluorescein dextran
Manufactured by Thermo Fisher Scientific
Co-injected fluorescein dextran is a laboratory reagent used to facilitate the visualization and tracking of fluid flow and distribution in various experimental settings. It consists of fluorescein, a fluorescent dye, covalently linked to dextran, a polysaccharide. When co-injected with a sample, the fluorescent dextran enables researchers to observe the movement and dispersion of the fluid of interest.
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Lab products found in correlation
2 protocols using co injected fluorescein dextran
1
Lineage Tracing in Xenopus Embryos
2
Fluorescent Lineage Tracing and Immunofluorescence in Xenopus Embryos
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Co-injected fluorescein dextran (70,000 MW, ThermoFisher, D1822) was used as lineage tracer for dorsal lips in IF analyses. For IF analyses, embryos were fixed in 4% paraformaldehyde for 1 h at RT, followed by 2 washes in 1x PBS-for 15 min each. For staining of animal caps or bisected specimens, embryos were manually dissected horizontally or sagittaly after fixation, transferred to 24 well plates, and washed twice for 15 min in PBST (PBS/0.1% Triton X-100). After blocking for 2h at RT in CAS-Block (1:10 in PBST; ThermoFisher, #008120) blocking reagent was replaced by antibody solution (diluted in CAS-Block) for incubation ON at 4°C. Antibodies used were: β1integrin (DSHB 8C8-s; 1:70), Ctnnb1 (clone H102; Santa Cruz Biotechnology sc-7199; 1:200), 1:200) . Then antibody solution was removed and explants washed twice for 15 min in PBS. Secondary antibodies (ThermoFisher, all 1:1000 in CAS-Block) were incubated for 2 h at RT. Cell borders were visualized using AlexaFluor TM Plus 405 Phalloidin (ThermoFisher A30104) overnight at 4°C (1:100 in PBS-). For photo documentation, bisected embryos or caps were transferred onto microscope slides or positioned in low melt agarose on a Petri dish (0.5% low melt agarose in 1x PBS-).
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