Table 5.

Reagent calculation for whole blood stimulation and reference sample preparation

	Donor		Reference sample
	1 tube	4 experiments +10%	1 tube		4 experiments + 10%
Stimulation agent 2×/plain medium	500 μL	2,200 μL	250 μL (RSQ)	250 μL (RPMI)	1100 μL (RSQ)	1100 μL (RPMI)
Blood	500 μL	2,200 μL	250 μL	250 μL	1100 μL	1100 μL
Protein transport inhibitor	2 μL	8.8 μL	1 μL	1 μL	4.4 μL	4.4 μL
CisPt (5 mM)	1 μL	4.4 μL	1 μL		4.4 μL
Proteomic Stabilizer	1.4 mL	6.2 mL	1.4 μL		6.2 mL

Reagent volumes needed to stimulate, fix and freeze 1 donor sample in 4 experiments (columns for Donor) and 1 reference sample in 4 experiments (columns for reference sample). Note that half of the reference sample is stimulated with RSQ and the other half is left unstimulated. These samples are pooled after stimulation; thus the calculation is done separately up to the protein transport inhibitor.