F10-Ham's

for all LLCPk cell lines

Thaw FBS and anti-anti first!

To make	mL F10-Ham's
start with	mL water
add:	g F10 (Hams) (Sigma N6635)
add:	g Optimem (Invitrogen 226000-050)
add:	g NaHCO3
add:	g HEPES, then pH to 7.2 (initial pH = ~7.1)
add:	mL FBS
add:	mL anti/anti

bring to final volume, filter sterilize in BSC, refrigerate

Ingredient	supplier	cat #	1 L	500 mL	250 mL
F10 (Ham's)	Sigma	N6635	4.9	2.45	1.225	g
Optimem	Invitrogen	22600-050	6.8	3.4	1.7	g
NaHCO3			1.8	0.9	0.45	g
HEPES			0.66	0.33	0.165	g
FBS	Krackeler	12103C	75	37.5	18.75	mL
anti/anti	Fisher	SV30079.01	10	5	2.5	mL

Mix F10, Optimem, NaHCO3, HEPES in about 70% of the final volume of dH2O.

Initial pH ~7.1
Adjust pH to 7.2 (it will rise to 7.3 in the CO2 incubator).
Add appropriate aliquot of FBS and antibiotic/antimycotic.
Bring to final volume.
Filter sterilize in the tissue culture hood.
Refrigerate

For serum free, replace serum with distilled water