Embryos in the morpholino validation experiment were fixed in 3% PFA in PBS for 2 hours, washed in PBST (1X PBS and 0.1% Triton X-100) and then stained with phallodin-647 (Invitrogen). Embryos used for centriole analysis were fixed in 100% ice-cold methanol for 48 hours at −20°C. Embryos were rehydrated in a methanol series, washed in PBST, and blocked in 10% heat-inactivated goat serum for 2 hours. Mouse Anti-centrin (EMD Millipore 04–1624) and rabbit anti–ZO-1 (61–7300; Invitrogen) primary antibodies were used, followed by Cy-2 and Cy-3-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.).
Phallodin 647
Manufactured by Thermo Fisher Scientific
Phallodin-647 is a fluorescent dye used in cell biology and microscopy applications. It selectively binds to filamentous actin (F-actin) within cells, allowing for the visualization and analysis of the cytoskeleton.
Automatically generated - may contain errors
2 protocols using phallodin 647
1
Immunostaining of Embryonic Structures
2
Immunostaining of Embryonic Structures
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Embryos in the morpholino validation experiment were fixed in 3% PFA in PBS for 2 hours, washed in PBST (1X PBS and 0.1% Triton X-100) and then stained with phallodin-647 (Invitrogen). Embryos used for centriole analysis were fixed in 100% ice-cold methanol for 48 hours at −20°C. Embryos were rehydrated in a methanol series, washed in PBST, and blocked in 10% heat-inactivated goat serum for 2 hours. Mouse Anti-centrin (EMD Millipore 04–1624) and rabbit anti–ZO-1 (61–7300; Invitrogen) primary antibodies were used, followed by Cy-2 and Cy-3-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.).
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