Difference between revisions of "Rapamycin Injections"
From Bridges Lab Protocols
(→Preparation) |
(added SOPs) |
||
| (6 intermediate revisions by 3 users not shown) | |||
| Line 1: | Line 1: | ||
| + | ==SOP== | ||
| + | *[[Safety and Animal Training]] | ||
| + | *[[SOP - Needle Capping]] | ||
| + | *[[SOP - Rapamycin]] | ||
| + | |||
==Materials== | ==Materials== | ||
*Tween 80 - Sigma P1754 | *Tween 80 - Sigma P1754 | ||
| − | *PEG - Sigma P3265 | + | *PEG 400 Liquid - Sigma P3265 |
==Preparation== | ==Preparation== | ||
| − | * | + | *Weigh out 2.08g of each Tween 80 and PEG into a 50 mL falcon tube. Bring up to 40 mL with water and filter by steriflip. This is 5.2% PEG/Tween |
| − | *Dissolve rapamycin at 20 mg/mL in | + | *Dissolve rapamycin at 20 mg/mL (0.0219 M) in ethanol (prepare as a 1 mL aliquot and store at -20 C). |
| − | *Add | + | *Add 5 uL of 20 mg/mL RAPA in 1 mL Tween 80/PEG diluent to get a 0.1 mg/mL concentration (109 uM; use this for 1 week). |
| + | *IP inject 10 uL/g body weight (1 mg/kg) | ||
| + | |||
| + | ==Alternate Rapamycin Injection Regimen== | ||
| + | * Phenotypes seem to be different with rapamycin as treated by Lamming et al. | ||
| + | * This protocol is from Lamming et al http://dx.doi.org/10.1126/science.1215135 | ||
| + | <blockquote> | ||
| + | rapamycin treatment was performed by injecting 8 to 10 week old mice intraperitoneally | ||
| + | once daily with either 1 mg/kg rapamycin suspended in 0.9% NaCl and 2% ethanol at a | ||
| + | concentration of 0.5 mg/mL (547 μM), or vehicle only for 14-28 days. | ||
| + | </blockquote> | ||
Latest revision as of 17:07, 8 June 2020
SOP
Materials
- Tween 80 - Sigma P1754
- PEG 400 Liquid - Sigma P3265
Preparation
- Weigh out 2.08g of each Tween 80 and PEG into a 50 mL falcon tube. Bring up to 40 mL with water and filter by steriflip. This is 5.2% PEG/Tween
- Dissolve rapamycin at 20 mg/mL (0.0219 M) in ethanol (prepare as a 1 mL aliquot and store at -20 C).
- Add 5 uL of 20 mg/mL RAPA in 1 mL Tween 80/PEG diluent to get a 0.1 mg/mL concentration (109 uM; use this for 1 week).
- IP inject 10 uL/g body weight (1 mg/kg)
Alternate Rapamycin Injection Regimen
- Phenotypes seem to be different with rapamycin as treated by Lamming et al.
- This protocol is from Lamming et al http://dx.doi.org/10.1126/science.1215135
rapamycin treatment was performed by injecting 8 to 10 week old mice intraperitoneally once daily with either 1 mg/kg rapamycin suspended in 0.9% NaCl and 2% ethanol at a concentration of 0.5 mg/mL (547 μM), or vehicle only for 14-28 days.
