. 2020 Jun 20;10(12):e3658. doi: 10.21769/BioProtoc.3658

Table 1. Stock solutions and concentration for the preparation of Kinase buffer.

Reagent	Final concentration in reaction tube (30 µl)	Stock concentration	Concentration in 3x Kinase buffer	Vol of stock for 3x Kinase buffer (µl)		Notes
Distilled water	-	-		24.85		-
HEPES	25 mM	200 mM, pH 7.3	75 mM	131.25		-
G3P	40 mM	800 mM	120 mM	52.5		-
BSA	1 mg/ml	50 mg/ml	3 mg/ml	21		-
MgCl₂	10 mM	1 M	30 mM	10.5		-
Na₃VO₄	100 µM	100 mM	300 µM	1.05		-
NPP	5 mM	500 mM	15 mM	10.5		-
IBMX	0.1 mM	100 mM (in DMSO)	0.3 mM	1.05		-
cAMP	1 mM	1 M	3 mM	1.05		-
ATP	40 µM	1 mM	120 µM	42		*Add just before use
*Protease inhibitor	1x	25x	2x	28
*Kemptide	100 µM	5 mM	300 µM	21
**ATP-³²Pγ	1 µCi/assay	10 µCi/µl	3 µCi/assay	5.25		**Add in the radioisotope room
Total Volume					350	-