List of Buffers
Carbonate Buffer for coating ELISA
0.1 M Na2CO3 adjust pH to 9.6 with 0.1 M NaH2CO3
Coagulation buffer
A drop of EDTA 100mM in an eppendorf tube that will receive the blood is enough.
Complete cutlture RPMI Medium
Gibco RPMI 1640 with L-Glutamine (Ref 21875) –>500ml
All from Gibco:
+ Glutamine 2mM (if the medium do not contain any) –> 5ml
+ Sodium Pyruvate 100mM (Ref 11360) (100x) –> 5ml
+ MEM Non Essential Aminos Acids (Ref 11140) (100x) –> 5ml
+ Hepes 1M (Ref 15630) (100x) –> 5ml
+ Peni-Strep (Ref 15140) (100x) –> 5ml
+ Gentamycin 50mg/ml (Ref 15750) (1000x) –> 500ul
+ βMercaptoethanol 50mM (Ref 31350) (1000x) –> 500ul
+ FCS 10% –> 50ml
Confocal Buffer
PBS, 10%, FCS, 0.1%Azide
DABCO (immunomount pour slide pour confocal)
90% glycerol
10% PBS
2.5g/100ml DABCO [1,4-Diazabicyclo(2,2,2)octane]
Dissolve de DABCO first in PBS (azide at 0.1/0.05%), then add the glycerol little by little while shaking.
Adjust the pH to 8.6 by adding 2NHCl ~ 1 to 2ml (or 800 μl of 12N HCl in 100ml).
FACS Buffer
PBS, 0.1% (w/v) sodium azide, 1% FCS, EDTA 2mM
Immunohistochemistry Buffer – Tris pH7.6
1 part of TRIS 0.2M (121.14g / 5L)
1.5 part of NaCl (42.5g / 5L)
1.5 part of 0.1N HCl (50ml / 5L)
T cell in vitro stimulation
PMA (2. Phorbol 12-Myristate 13-Acetate) 50ng/ml and Ionomycin 1μg/ml
4h for cytokines intracellular staining; 24h for cytokines ELISA
List of Buffers —> PDF