Differentiation of 3T3-L1 Cells: Difference between revisions
updated MIX stock solution instructions |
added SOP requirements |
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__NOTOC__ | __NOTOC__ | ||
==SOP== | |||
*[[SOP- Biosafety Cabinets]] | |||
*[[Sop- Compressed Gases]] | |||
*[[SOP- Vacuum Pumps]] | |||
==Materials== | ==Materials== | ||
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*'''Insulin''' (Sigma I-5523). Dissolve in 0.01N Hydrochloric acid (28.7μl/50ml) as a 1 mg/ml stock. Aliquot into 1.5 mL tubes and store at -20 | *'''Insulin''' (Sigma I-5523). Dissolve in 0.01N Hydrochloric acid (28.7μl/50ml) as a 1 mg/ml stock. Aliquot into 1.5 mL tubes and store at -20 | ||
*'''Dexamethasone''' (Sigma D-1756). Dissolve in ethanol as a 2.5 mM stock (0.049g/50ml). Aliquot into 1.5 tubes and store at -20. | *'''Dexamethasone''' (Sigma D-1756). Dissolve in ethanol as a 2.5 mM stock (0.049g/50ml). Aliquot into 1.5 tubes and store at -20. | ||
*'''MIX''' (Sigma I-5879). Add 2.78g MIX and 0.98g KOH and bring up to 50 mL. Aliquot into 1.5 mL tubes and store at -20 | *'''MIX''' (Sigma I-5879). Add 2.78g MIX and 0.98g KOH and bring up to 50 mL (250 mM stock). Aliquot into 1.5 mL tubes and store at -20 | ||
*'''Fibroblast Media''' (DMEM + 10% NCS + 1X PSG; Sterile filter into new bottle) | *'''Fibroblast Media''' (DMEM + 10% NCS + 1X PSG; Sterile filter into new bottle) | ||
*'''Adipocyte Media''' (DMEM + 10 % L1-FBS + 1X PSG; Sterile filter into new bottle) | *'''Adipocyte Media''' (DMEM + 10 % L1-FBS + 1X PSG; Sterile filter into new bottle) | ||