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. 2016 Feb;38:56–67. doi: 10.1016/j.neurobiolaging.2015.10.021

Table 2.

Reagents and sample preparation of Neuroflammation Panel 1 (human) kit from MSD multispot assay system

MSD panel kits Analysts Diluent for sample and calibrator Dilution Added volume for diluent (μL) Sample (μL) Diluent for detection antibody
Pronflammatory panel 1 IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-13, TNF-α Diluent 2 1:2 25 25 Diluent 3
Cytokine panel 1 IL-1α, IL-5, IL-7, IL-12p40, IL-15, IL-16, IL-17A, TNF-β, VEGF Diluent 43 1:2 25 25 Diluent 3
Chemokine panel 1 Eotaxin, MIP-1β, Eotaxin-3, TARC, IP-10, MIP-1α, MCP-1, MDC, MCP-4 Diluent 43 1:4 37 13 Diluent 3
Angiogenesis panel 1 VEGF-C, VEGF-D, Tie-2, Flt-1, PIGF Diluent 7 1:2 25 25 Diluent 11
Vascular injury panel 1 SAA, CRP, VCAM-1, ICAM-1a Diluent 101 1:5 20 5 Diluent 101
Human bFGF Kit V-PLEX bFGFb Diluent 7 1:2 25 25 Diluent 11

Key: CRP, C-reactive protein; FGF, fibroblast growth factor; Flt-1, also known as VEGF-1 receptor; ICAM, intercellular adhesion molecule; IFN, interferon; IL, interleukin; IP, interferon-inducible protein; MCP, monocyte chemoattractant protein; MDC, macrophage-derived chemokine; MIP, macrophage inflammatory protein; PIGF, placenta growth factor; SAA, serum amyloid A; TARC, thymus and activation-regulated chemokine; TNF, tumor necrosis factor; VCAM, vascular cell adhesion molecule; VEGF, vascular endothelial growth factor.

a

For vascular injury, prepare 50 μL (40 μL of diluent 101 and 10 μL of normalized sample) in the transfer plate and add 25 μL into the kit plate.

b

The concentration of bFGF was extremely high in the all brain tissue. A single human bFGF kit had to be used for this specific cytokine, and samples were diluted 1:100 with lysis buffer and then diluted 1:2 with Diluent 7.