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20-hydroxyecdysone (20E) feeding of Drosophila larvae

JJ Jacob S Jaszczak
LD Laura DeVault
LJ Lily Jan
YJ Yuh Nung Jan

Last updated date: Dec 28, 2022 Views: 680 Forks: 0

original An abbreviated version of this protocol was published in eLife in Mar, 2022
Steroid hormone signaling activates thermal nociception during Drosophila peripheral nervous system development
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Sigma Aldrich

20-Hydroxyecdysone

≥93% (HPLC), powder

Catalog number: H5142

CAS Number: 5289-74-7

 

  1. Create a 50uM working stock with powder 20-Hydroxyecdysone in 100% ethanol. Aliquot the working stock and store at -20oC. 
  2. Mix the 20E working stock with room temperature cornmeal-molasses media to create the desired concentration of supplemented media (mg 20E / ml media). 1 ul working stock = 0.024mg 20E. Stir media for 90 secs to fully distribute the 20E into the food. 
  3. Create control media by mixing a volume of ethanol equivalent to the experimental group with cornmeal-molasses media (ul EtOH / ml media). Stir media for 90 secs to fully distribute the ethanol into the food. 
  4. Let the media mixture sit for 30 minutes at room temperature to allow the ethanol to evaporate before adding larvae to the media.
  5. Transfer groups of larvae to supplemented media and allow larvae to feed. The dosage of 20E to larvae is dependent on ingestion of supplemented media. 

 

 

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Jaszczak, J, DeVault, L, Jan, L and Jan, Y(2022). 20-hydroxyecdysone (20E) feeding of Drosophila larvae. Bio-protocol Preprint. bio-protocol.org/prep2097.
  2. Jaszczak, J. S., DeVault, L., Jan, L. Y. and Jan, Y. N.(2022). Steroid hormone signaling activates thermal nociception during Drosophila peripheral nervous system development. eLife. DOI: 10.7554/eLife.76464

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