Difference between revisions of "Preparation of High Serum for A1 cells"
Jump to navigation
Jump to search
(Created page with "For 400 ml FCS 40 ml Eagle’s MEM 252 ml DdH2O 100 ml Pen/ Strep 4 ml Glutamin 4 ml Insulin (stock: 1mg/ ml) 4 ml - grow at 25ºC in 2% CO2 - if you want to gr...") |
|||
Line 1: | Line 1: | ||
− | For 400 ml | + | [[For 400 ml]] |
FCS 40 ml | FCS 40 ml |
Revision as of 10:09, 13 February 2015
FCS 40 ml Eagle’s MEM 252 ml DdH2O 100 ml Pen/ Strep 4 ml Glutamin 4 ml Insulin (stock: 1mg/ ml) 4 ml
- grow at 25ºC in 2% CO2 - if you want to grow without CO2, then substitute L-15 for the MEM
- passage once per week 1:3
- wash with APBS (100 ml PBS + 25 ml ddH2O) - trypsinize with TE - stop with media - centrifuge cells at 100 rpm 3 minutes
to make myotubes:
- wash cells 1x with APBS - put cells in 0.5% FCSmedia - wait 3 days