Micronized dHACM (n = 4–5) was incubated in lysis buffer (RayBiotech) containing Protease Inhibitor Cocktail Set III at a concentration of 35 mg/mL with gentle agitation overnight at 4°C. Lysates were centrifuged and filtered (0.22 μm pore size; EMD Millipore). Growth factors, cytokines, proteases, and protease inhibitors were evaluated by using multiplex ELISA Quantibody® arrays (Human Cytokine Array Q1000 and Human MMP Array Q1, RayBiotech), and single-factor ELISAs (alpha 2-macroglobulin [α2 M] and alpha-1 antitrypsin [A1AT]; RayBiotech). Amounts of each molecule were reported as average moles/mg of tissue.
Human cytokine array q1000
Manufactured by RayBiotech
The Human Cytokine Array Q1000 is a multiplex immunoassay panel that allows for the simultaneous detection and quantification of 1,000 different human cytokines, chemokines, and growth factors in a single experiment. The array utilizes antibody-based capture technology to provide a comprehensive analysis of the human immune system's signaling molecules.
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2 protocols using human cytokine array q1000
1
Multiplex Evaluation of Micronized dHACM
2
Secretome Profiling of Adipose-Derived Stem Cells
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Conditioned media was prepared by cultivation of ADSC in MEM α supplemented without FCS for 24 h. Afterwards, the media was concentrated and screened for 80 human inflammatory and growth factors (BLC, Eotaxin‐1, Eotaxin‐2, G‐CSF, GM‐CSF, I‐309, ICAM‐1, IFN‐gamma, IL‐1 alpha, IL‐1 beta, IL‐1 ra, IL‐2, IL‐4, IL‐5, IL‐6, IL‐6 sR, IL‐7, IL‐8, IL‐10, IL‐11, IL‐12 p40, IL‐12 p70, IL‐13, IL‐15, IL‐16, IL‐17, MCP‐1, M‐CSF, MIG, MIP‐1 alpha, MIP‐1 beta, MIP‐1 delta, PDGF‐BB, RANTES, TIMP‐1, TIMP‐2, TNF alpha, TNF beta, sTNFRI, sTNFRII Amphiregulin, BDNF, bFGF, BMP‐4, BMP‐5, BMP‐7, b‐NGF, EGF, EGFR, EG‐VEGF, FGF‐4, FGF‐7, GDF‐15, GDNF, Growth Hormone, HB‐EGF, HGF, IGFBP‐1, IGFBP‐2, IGFBP‐3, IGFBP‐4, IGFBP‐6, IGF‐1, Insulin, M‐CSF R, NGF R, NT‐3, NT‐4, Osteoprotegerin, PDGF‐AA, PLGF, SCF, SCF R, TGF alpha, TGF beta 1, TGF beta 3, VEGF‐A, VEGFR2, VEGFR3, VEGF‐D) with a multiplex ELISA antibody array (Human Cytokine Array Q1000, RayBiotech Life Inc.).
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