Source of H10 hybridoma : available from RIKEN Cell Bank, Tsukuba, Japan

Solutions :

500 ml IMDM cell culture medium (Bioproducts, ref. M12722F) supplemented with 25mM Hepes (Bio Whittaker)

Fœtal calf serum (FCS)
100 ml L-glutamine 200mM (Gibco, ref. 250 30 024)

100 ml antibiotiques solution :10,000 U penicilline (Gibco, ref. 15 140114) + 10,000 U streptomycine

100ml Na pyruvate (Gibco, ref. 11360 039)

mercapto-ethanol solution : dilute 71 µl of mercaptoethanol (14 M ; Roth) in10 ml H₂O : filter on 0.22 µm Millex

For 100 ml of cell culture medium :

• 10 ml FCS
• 2 ml L glutamine
• 2 ml Na pyruvate
• 1 ml antibiotics
• 50 µl mercapto-ethanol

Protocol description : dilute the hybridoma cells at a final cell density of 2 to 5 x 10⁵ cells/ml

Reference :

Kawamitsu, H. et al. (1984) Biochemistry, 23, 3771-7 Monoclonal antibodies to poly(adenosine diphosphate ribose) recognize different structures.