R 10 (medium for KY1 cells)

Reagent	Final Conc.	Volume	Mass
dH₂O		800 ml
RPMI 1640 powdered media (Sigma R-6504)			1 bottle
NaHCO₃			2.2 g
L-glutamine 200 mM solution (Sigma G7513)	2 mM	10 ml
2-mercaptoethanol (MW 78.13, 14.2 M, Biorad 161-0710)	0.1 mM	7 µl

adjust pH to 7.2,
sterile filter, add 10 ml pen/strep (100x) to a final concentration of 100 U/ml penicillin, 100 µg/ml streptomycin,
add 10 % FBS
add IL-2 stock solution (10⁵ U/ml) directly to cells (start with 10³ U/ml, passing a few times before gradually weaning down to 50 U/ml, which is adequate for normal maintenance)
Freeze 100 x (200 mM) L-Glutamine solution in 10 ml aliquots

HEPES (MW 260.3)

10 mM

2.6 g

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