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V plex human proinflammatory panel

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The V-PLEX Human Proinflammatory Panel is a multiplex assay that enables the simultaneous quantification of multiple human proinflammatory biomarkers in a single sample. The panel is designed to measure the concentrations of specific proteins related to inflammatory processes.

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Lab products found in correlation

Human vascular injury 1 kit, by Mesoscale (2 mentions) Meso quick plex sq 120 mmreader, by Mesoscale (1 mentions) Human baff quantikine elisa, by R&D Systems (1 mentions) Quickplex sq 120, by Mesoscale (1 mentions)

7 protocols using v plex human proinflammatory panel

1

Biomarker Dynamics in ECMO Patients

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Blood samples were obtained daily during ECMO from the ECMO circuit or existing indwelling catheters, at the same time as routine clinical blood draws. Plasma was separated by centrifugation and stored at −80°C. Plasma concentrations for interferon gamma (IFNγ), interleukin-6 (IL-6), interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNFα), E-selectin, P-selectin, intercellular adhesion molecule-3 (ICAM-3), thrombomodulin (TM), tissue plasminogen activator (tPA), and plasminogen activator inhibitor-1 (PAI-1), were measured for ECMO days 1, 2, 3 and 5. Measurements for ECMO day 7 and every third day thereafter until ECMO decannulation were conducted if ECMO duration was longer than 5 days.
All assays were conducted in the Johns Hopkins Institute for Clinical and Translational Research (ICTR) Clinical Research Unit: Core Laboratory, using the Meso Scale Discovery (Rockville, MD) V-PLEX Human Proinflammatory Panel (for IFNγ, IL-6, IL-1β, TNFα), the Human Vascular Injury I Kit (for E-selectin, P-selectin, ICAM-3, TM), and single plex assays for tPA and PAI-1. Dynamic ranges for each analyte have been published.21 -23 The inter-assay coefficients of variation for the two multiplex assays used were: IFNγ, 7.96; IL-6, 15.93; IL-1β, 0.02; TNFα, 5.21; E-selectin, 6.45; P-selectin, 14.39; ICAM-3, 11.78; TM, 8.6. Laboratory personnel were blinded to clinical data.
Caprarola S.D., Ng D.K., Carroll M.K., Tekes A., Felling R.J., Salorio C.F., Almuqati R., Schwartz J.M., Everett A.D, & Bembea M.M. (2021). Pediatric ECMO: Unfavorable Outcomes are Associated with Inflammation and Endothelial Activation. Pediatric research, 92(2), 549-556.
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2

Measuring Inflammatory Cytokines in Synovial Fluid

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Synovial fluid was aspirated 3 or 4 days after surgery and was stored at –80°C
until measurement. Inflammatory cytokine levels in the synovial fluid were
measured using a multiplex enzyme-linked immunosorbent assay system (V-PLEX
human proinflammatory panel; Meso Scale Discovery) and MESO Quick Plex SQ 120 MM
Reader (Meso Scale Discovery). IL-1β, IL-2, IL-6, IL-8, IL-10, TNF-α, and IFN-γ
levels were quantified according to the manufacturer’s protocol. Nondiluted
synovial fluid was transferred into 96-well plates and then measured.
Before surgery and at 3 or 4 days after surgery, the C-reactive protein (CRP)
level and erythrocyte sedimentation rate (ESR) in the blood were measured.
Nakagawa Y., Tsuji K., Nakamura T., Katagiri H., Ozeki N., Shioda M., An J.S., Yoshida R., Sekiya I, & Koga H. (2023). Association of Infrapatellar Fat Pad Fibrosis at 3 Months After ACL Reconstruction With Short-term Clinical Outcomes and Inflammatory Cytokine Levels in the Synovial Fluid. Orthopaedic Journal of Sports Medicine, 11(4), 23259671231164122.
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3

Quantification of Immune Biomarkers

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Plasma samples collected were stored at −80°C. Circulating levels of B cell activation factor (BAFF) were measured using a human BAFF Quantikine ELISA (R&D Systems). Cytokine plasma concentrations were quantified by electrochemiluminescence immunoassay using the V-Plex human Proinflammatory panel (Meso Scale Discovery). All reagents and standards were provided by each manufacturer. Samples and standards were prepared according to each manufacturer’s instructions.
Lagou V., Garcia-Perez J.E., Smets I., Van Horebeek L., Vandebergh M., Chen L., Mallants K., Prezzemolo T., Hilven K., Humblet-Baron S., Moisse M., Van Damme P., Boeckxstaens G., Bowness P., Dubois B., Dooley J., Liston A, & Goris A. (2018). Genetic Architecture of Adaptive Immune System Identifies Key Immune Regulators. Cell Reports, 25(3), 798-810.e6.
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4

Systemic Cytokines as CRPS Risk Indicator

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Systemic levels of proinflammatory cytokines were assessed using a Meso Scale Diagnostics V-Plex Human Proinflammatory Panel employing electrochemiluminescent detection (Meso Scale Diagnostics, Rockville, MD). The current study focused on systemic levels of TNF-a as a biological indicator of pre-operative proinflammatory status and potential risk mechanism for CRPS41 (link),44 (link),46 (link),62 (link),81 (link). Plasma samples (described below) were analyzed in duplicate and assays were performed per manufacturer protocol and read and analyzed on a QuickPlex SQ120 (Meso Scale Diagnostics, Rockville, MD). Assay results are expressed as the mean of these duplicate assays (in pg/mL). The decision to obtain blood samples for cytokine assays was not made until after the study was ongoing, and these samples were therefore available for only n=72 participants.
Bruehl S., Billings FT I.V., Anderson S., Polkowski G., Shinar A., Schildcrout J., Shi Y., Milne G., Dematteo A., Mishra P, & Harden R.N. (2022). Preoperative Predictors of Complex Regional Pain Syndrome Outcomes in the 6 Months Following Total Knee Arthroplasty. The journal of pain, 23(10), 1712-1723.
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5

Biomarker Dynamics in ECMO Patients

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Blood samples were obtained daily during ECMO from the ECMO circuit or existing indwelling catheters, at the same time as routine clinical blood draws. Plasma was separated by centrifugation and stored at −80°C. Plasma concentrations for interferon gamma (IFNγ), interleukin-6 (IL-6), interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNFα), E-selectin, P-selectin, intercellular adhesion molecule-3 (ICAM-3), thrombomodulin (TM), tissue plasminogen activator (tPA), and plasminogen activator inhibitor-1 (PAI-1), were measured for ECMO days 1, 2, 3 and 5. Measurements for ECMO day 7 and every third day thereafter until ECMO decannulation were conducted if ECMO duration was longer than 5 days.
All assays were conducted in the Johns Hopkins Institute for Clinical and Translational Research (ICTR) Clinical Research Unit: Core Laboratory, using the Meso Scale Discovery (Rockville, MD) V-PLEX Human Proinflammatory Panel (for IFNγ, IL-6, IL-1β, TNFα), the Human Vascular Injury I Kit (for E-selectin, P-selectin, ICAM-3, TM), and single plex assays for tPA and PAI-1. Dynamic ranges for each analyte have been published.21 -23 The inter-assay coefficients of variation for the two multiplex assays used were: IFNγ, 7.96; IL-6, 15.93; IL-1β, 0.02; TNFα, 5.21; E-selectin, 6.45; P-selectin, 14.39; ICAM-3, 11.78; TM, 8.6. Laboratory personnel were blinded to clinical data.
Caprarola S.D., Ng D.K., Carroll M.K., Tekes A., Felling R.J., Salorio C.F., Almuqati R., Schwartz J.M., Everett A.D, & Bembea M.M. (2021). Pediatric ECMO: Unfavorable Outcomes are Associated with Inflammation and Endothelial Activation. Pediatric research, 92(2), 549-556.
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6

Multiplex ELISA Cytokine Quantification

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Cytokine levels in synovial fluid were quantitated by multiplex ELISA system (V-PLEX human pro-inflammatory panel, Meso Scale Discovery, MD). Levels of TNF-alpha, IL1-beta, IL2, IL6, IL8, IL10, and IFN-gamma in synovial fluid were quantitated according to the manufacturer’s protocol.
Inoue M., Muneta T., Ojima M., Nakamura K., Koga H., Sekiya I., Okazaki M, & Tsuji K. (2016). Inflammatory cytokine levels in synovial fluid 3, 4 days postoperatively and its correlation with early-phase functional recovery after anterior cruciate ligament reconstruction: a cohort study. Journal of Experimental Orthopaedics, 3, 30.
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7

Quantifying Inflammatory Cytokine Profiles

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IL-8, IL-6, IL-1β, TNF-α, and IFN-γ in cell-free supernatants collected from
neutrophils after culture at 5 × 105 cells/well in 96-well tissue
culture plates with E. coli lipid A or medium for 20 h were
quantified by electrochemiluminescence assay using the V-PLEX Human
Proinflammatory Panel (Meso Scale Discovery). GM-CSF was assayed by colorimetric
ELISA (Human GM-CSF ELISA Ready-SET-Go kit, eBioscience; limit of detection 10
pg/ml).
SenGupta S., Rane M.J., Uriarte S.M., Woolley C, & Mitchell T.C. (2019). Human neutrophils depend on extrinsic factors produced by monocytes for their survival response to TLR4 stimulation. Innate Immunity, 25(8), 473-486.
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