Table 49.

Buffers and Kits

Buffer/Kit
PBS/EDTA	PBS supplemented with 2 mM EDTA
flow cytometry buffer	PBS supplemented with 2 mM EDTA and 0,2% FCS
40% Percoll	prepare an isotonic 90% Percoll buffer using 10× PBS and dilute to 40% in flow cytometry buffer
67% Percoll	prepare an isotonic 90% Percoll buffer using 10× PBS and dilute to 67% in flow cytometry buffer
DTT buffer	25 ml HBSS Ca2+ and Mg2+ free 25 ml 10× HEPES Bicarbonat Buffer (see below) 25 ml FBS 175 ml Aq dest Store at room temperature, refresh every 2 weeks Freshly add 7,7 mg DTE/ 50 ml buffer. Use 10 ml/sample
Digestion media	500 ml RPMI 1640 1% FBS 1× HGPG (see below) 1 mM CaCl2 1 mM MgCl2 Freshly add 0,1 mg Collagenase D and 0,1 mg DNaseI/ml. Use 6 ml/sample
10× HEPES Bicarbonate Buffer	23,8 g HEPES 21 g Sodium Bicarbonat 1 L distilled water Adjust pH7,2 with HCl
100x HGPG	14,9 g HEPES 1×10^6 U Penicillin 2,5 mg Gentamycin 125 ml RPMI+Glutamax Adjust to pH7,5, filter sterile Store at −20°C
ACK lysing buffer	ThermoFisherScientific Cat#A10492-01
Clicks full medium	500 ml Clicks medium (IrvineScientific, 9195) 10% FCS (Gibco, 10500-064) 5 ml GlutaMAX Supplement (Gibco, 13462629) 5 ml Penicillin/Streptomycin (Gibco, 15140-122)
Streptavidin Microbeads	Miltenyi Biotec 130-048-101
CD4 (L3T4) Microbeads, mouse	Miltenyi Biotec 130-117-043