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.
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.
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.