Method/Kit:
Luminex Human Discovery Assay (8-Plex) LXSAHM-08

Measurement Type Fluorescence Intensity
Description The Luminex Assay is a bead-based immunoassay multiplex technique that allows multiple analytes
(ex. cytokines, chemokines, metabolic biomarkers, etc.) to be quantitated simultaneously in a
single biological matrix sample. Generally, the results obtained using a standardized Luminex
assay are superior or equivalent to conventional ELISA, even as the differentiated xMAP
technology allows high throughput biomarker testing with reduced sample volume, test
preparation, and reagent costs. Luminex immunoassays can multiplex up to 100 compatible
analytes simultaneously using a commercial or custom kit. The 8-Plex quantitates 8 analytes simultaneously and was used for the hToc experiment at CMSR @ UR. CMSR @ UR chose the 8 cytokines (Monocyte Chemoattractant Protein-1, Interleukin 6, C-C Motif Chemokine Ligand 3, Interleukin 10,
C-X-C Motif Chemokine Ligand 10, Interleukin 1 beta, Tumor necrosis Factor alpha, Interleukin 17) to test the cell supernatant of our top well at different time points and culture conditions.

Target/Analytes with this Method/Kit
Target/Analyte Short Name Description
CCL3 CCL3 CCL3/MIP-1 alpha
CXCL10 CXCL10 CXCL10/IP-10/CRG-2
IL-10 IL-10 Senescence Associated Secretory Phenotype (SASP) protein
IL-17 IL-17 IL-17/IL-17A
IL-1B IL-1B Interleukin 1B
IL-6 IL 6 Interleukin 6
MCP-1 MCP-1 Monocyte Chemoattractant Protein-1, is a key chemokine that regulates migration and infiltration of monocytes/macrophages.
M/T D1 / MCP-1 M/T D1 / MCP-1 The M/TC D1 treatment group is comprised of tenocytes and M0 monocytes co-cultured in the collagen hydrogel for 7 days total, with measurement taken on day 1. No top component is attached. The MCP-1 cytokine subjected to this treatment is a key chemokine that regulates migration and infiltration of monocytes and macrophage
M/T D4 / MCP-1 M/T D4 / MCP-1 The M/TC D1 treatment group is comprised of tenocytes and M0 monocytes co-cultured in the collagen hydrogel for 7 days total, with measurement taken on day 4. No top component is attached. The MCP-1 cytokine subjected to this treatment is a key chemokine that regulates migration and infiltration of monocytes and macrophage
M/T D7 / MCP-1 M/T D7 / MCP-1 The M/TC D1 treatment group is comprised of tenocytes and M0 monocytes co-cultured in the collagen hydrogel for 7 days total, with measurement taken on day 7. No top component is attached. The MCP-1 cytokine subjected to this treatment is a key chemokine that regulates migration and infiltration of monocytes and macrophage
M/T/H / MCP-1 M/T/H / MCP-1 The M/TC D1 treatment group is comprised of tenocytes and M0 monocytes co-cultured in the collagen hydrogel for 7 days total, on day 8 endothelial cells are added to the system through the addition of the top component. The measurement is taken 72 hours post adding the endothelial cells. The MCP-1 cytokine subjected to this treatment is a key chemokine that regulates migration and infiltration of monocytes and macrophage
TNF Alpha TNF Alpha Tumor necrosis Factor alpha, is considered the “Master regulator” of the inflammatory cytokine production cascade.
TNF-Alpha TNFA Tumor necrosis factor (TNF, tumor necrosis factor alpha, TNFα, cachexin, or cachectin) is a cell signaling protein (cytokine) involved in systemic inflammation and is one of the cytokines that make up the acute phase reaction. It is produced chiefly by activated macrophages, although it can be produced by many other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons.
T-TGF-β1, SF / MCP-1 T-TGF-β1, SF / MCP-1 The TC-TGF-β1 treatment group is comprised of tenocytes in the collagen hydrogel cultured for 7 days with no exogenous treatment. No top component is attached. The MCP-1 cytokine subjected to this treatment is a key chemokine that regulates migration and infiltration of monocytes and macrophages.
T+TGF-β1, SF / MCP-1 T+TGF-β1, SF / MCP-1 The TC+TGF-β1 treatment group is comprised of tenocytes in the collagen hydrogel cultured for 7 days and exposed with TGF-β1 at 10 ng/mL. No top component is attached. The MCP-1 cytokine subjected to this treatment is a key chemokine that regulates migration and infiltration of monocytes and macrophages.

Studies Utilizing this Method/Kit
Study Start Date Study Types Description
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