Plasma was extracted from mouse peripheral blood samples collected on days 7, 14, and 21 of treatment (n = 8 per group at each time-point). Nine cytokines (interleukin-15, IL-15; interleukin-18, IL-18; basic fibroblast growth factor, bFGF; leukemia inhibitory factor, LIF; macrophage-colony stimulating factor, M-CSF; monokine induced by gamma interferon, MIG; macrophage inflammatory protein-2, MIP-2; platelet-derived growth factor-BB, PDGF-BB; and vascular endothelial growth factor, VEGF) known to be involved in angiogenesis were quantified in plasma using a Bio-Plex Pro mouse cytokine 9-plex assay and standard protocol (Bio-Rad Laboratories). To evaluate the integrity of endothelial and stromal cells, the uteri and ovaries of mice treated with ABT-898 or 5% dextrose were paraffin-embedded, sectioned (5 μm), and stained with vimentin primary antibody (1:500) (Ab7388; Abcam), cytokeratin primary antibody (1: 500) Z0662; DAKO)) and secondary polyclonal goat anti-rabbit biotinylated antibody (1:1000, (DAKO cytomation). The liver, kidney, ovary and uterus harvested from experimental mice on day 21 of treatment with ABT-898 or 5% dextrose were paraffin-embedded, sectioned (5 μm), and stained with hematoxylin and eosin and periodic acid schiff’s stain using a standard protocol.
Bio plex pro mouse cytokine 9 plex assay
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex Pro mouse cytokine 9-plex assay is a multiplex assay designed to simultaneously measure the concentration of nine different mouse cytokines in a single sample. The assay uses magnetic beads coated with antibodies specific to each cytokine, allowing for the quantification of multiple analytes in a small sample volume.
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3 protocols using bio plex pro mouse cytokine 9 plex assay
1
Cytokine Profiling and Tissue Histology
2
Cytokine Profiling in Cancer Cachexia
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Plasma samples were taken from cancer cachexia mice at 24 h after the evaluation of behavioral phenotypes. Cytokines, chemokines, and growth factors in plasma samples were quantified by the Bio-Plex® Multiplex System (Bio-Rad Laboratories Inc., CA, USA). In this study, the Bio-Plex Pro™ Mouse Cytokine 23-plex Assay (#64,140,214, Bio-Rad Laboratories Inc.) and Bio-Plex Pro™ Mouse Cytokine 9-plex Assay (#64,174,947, Bio-Rad Laboratories Inc.) were used.
3
Multiplex Cytokine Profiling in Hippocampus
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Cytokines and chemokines in lysates from hippocampus were detected with a cytometric bead array using a Bio-Plex Pro mouse cytokine 9-plex assay (Bio-Rad Laboratories, Hercules CA, United States) and analyzed with a Luminex 200 system (Bio-Rad Laboratories, Hercules CA, United States) according to the instructions of the manufacturer and as previously described (Lai et al., 2020 (link)).
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