Dechorionated embryos were cleared before light sheet imaging as previously described (Deryckere et al., 2020 (link)). Whole mount immunohistochemistry was performed for long term CFDA tracing, during the PBS washing steps in between incubation in ScaleCUBIC-1 and ScaleCUBIC-2 as follows: after clearing in ScaleCUBIC-1, embryos were washed four times; one time for 5 min and three times for 2 hr in PBS supplemented with 0.3% Triton X-100 (PBS-T) and then incubated in primary antibody solution in PBS-T (goat anti-fluorescein (Novus Biologicals NB600-493, 1:500, pre-incubated overnight with non-injected embryos)) for 2 days at 4°C. Embryos were then washed three times for 2 hr in PBS-T, secondary antibody solution was added (donkey anti-goat Alexa 488 (Life Technologies, 1:300)) and incubated overnight at 4°C after which the samples were washed three times for 2 hr in PBS-T and then incubated in 1/2-water diluted ScaleCUBIC-2 at 37°C.
Nb600 493
Manufactured by Novus Biologicals
NB600-493 is a lab equipment product manufactured by Novus Biologicals. It serves as a core function, but a detailed description without interpretation or extrapolation cannot be provided.
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Lab products found in correlation
2 protocols using nb600 493
1
Long-term CFDA tracing in cleared embryos
2
Immunohistochemistry in Cleared Embryos
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Dechorionated embryos were cleared before light sheet imaging as previously described (Deryckere et al., 2020) (link). Whole mount immunohistochemistry was performed for long term CFDA tracing, during the PBS washing steps in between incubation in ScaleCUBIC-1 and ScaleCUBIC-2 as follows: after clearing in ScaleCUBIC-1, embryos were washed 4 times; 1 time for 5 mins and 3 times for 2 hours in PBS supplemented with 0.3% Triton X-100 (PBS-T) and then incubated in primary antibody solution in PBS-T (goat anti-fluorescein (Novus Biologicals NB600-493, 1:500, pre-incubated overnight with noninjected embryos)) for 2 days at 4 °C. Embryos were then washed 3 times for 2 hours in PBS-T, secondary antibody solution was added (donkey anti-goat Alexa 488 (Life Technologies, 1:300)) and incubated overnight at 4 °C after which the samples were washed 3 times for 2 hours in PBS-T and then incubated in 1/2-water diluted ScaleCUBIC-2 at 37 °C.
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