View source for Hybridoma cells in SFX media

==Materials:==
<li>5 ml, 10 ml, 20 ml pipettes
<li>Serum free media( 400 mls SF media  + 1%   antibiotics 
                        4 mls P/S and 4 mls Glutamine
<li>2 blue cap flasks
<li>1 conical tube

==Execution:==
# fill 10 mls SFX media per blue cap flask
# put flask  for > = 1 hour in incubator 10% CO2 @ 37 °C  
# use 10 ml pipette to take up the media and blow off the cells 
# check the flask in microscope to make sure that the cells have detached
# use 10 ml pipette to remove the cells from the flask into the conical tube
# spin at 1000rpm for 3 minutes
# aspirate off the media
# add 2  mls  of SFX media
# resuspend the cells
# add the 2 mls of the suspension to 1flask
# ut in 10% CO2 incubator 
# let cells grow until they die ( 1-2 weeks; spin cells down( 1000 rpm , 3 min. 4°C)
# collect supernatent( contains Antibody)  and add  @1:50 concentratin 5 x sodium azide/ 500 mM Hepes and store at 4°C