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Preparation of TNFa
Contributor: Suprya Jayadev
Date: August 15, 1991
Krebs-Ringer-Hepes Buffer (w/ BSA):
NaCl 7.112 g
KCl 0.365 g
MgSO4 - 7 H2O 0.3 g
CaCl2 - 2 H2O 0.1 g
Hepes 2.86 g
--> adjust pH to 7.4
BSA* to 0.1%
--> readjust pH to 7.4
--> qs to 1 liter volume
--> filter sterilize
* = charcoal treated, fatty acid free BSA
Resuspend 1 mg of TNFa in 0.5 ml of Krebs-Ringer-Hepes Buffer containing
BSA
--> 117.6 µM solutin or
1 mg of TNFa resuspended in 0.588 ml of KRH/BSA buffer is a 100 µM
solution