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Mouse cytokine chemokine array 32 plex md 32

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The Mouse Cytokine/Chemokine Array 32-plex (MD-32) is a multiplexed assay that simultaneously measures the levels of 32 different mouse cytokines and chemokines in a single sample. It is designed to provide a comprehensive profile of the immune response in mouse models.

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Rneasy mini kit, by Qiagen (2 mentions) Rneasy fibrous tissue mini kit, by Qiagen (2 mentions)

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32-plex cytokine/chemokine array (4 citations) Mouse Cytokine/Chemokine Array 32-plex (2 citations)

5 protocols using mouse cytokine chemokine array 32 plex md 32

1

Investigating the Immunomodulatory Effects of IsC1ql3

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We determined the effects of IsC1ql3 on host cytokine production in vitro and in vivo. For in vitro assays, the murine splenocytes, CD4+ and CD8+ T cells were isolated as described above. The isolated cells were incubated with 1 μg/mL rIsC1ql3 or BSA (control) and stimulated by 10 ng/mL LPS or 106B. burgdorferi for 6 h. The cells and supernatant were harvested, and total RNA was extracted using a RNeasy Mini kit (Qiagen). For in vivo studies, tick IsC1ql3 gene was silenced as described above. Then the silenced nymphal ticks were put on mice ears. After 72 h post tick bite, mice were be sacrificed, and punch biopsy specimens were taken from the tick bite site. Total RNA was extracted, and cDNA was synthesized as described above. The qPCR primers sequences of cytokine genes are shown in Table S2. In addition, we also quantify cytokines production by the Mouse Cytokine/Chemokine Array 32-plex (MD-32) performed by Eve Technologies. Serum collected from each group of mice was sent for cytokine analyses. The cytokines represented by this array are Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, KC, LIF, LIX, MCP-1, M-CSF, MIG, MIP-1α, MIP-1β, MIP-2, RANTES, TNFα, and VEGF.
Tang X., Arora G., Matias J., Hart T., Cui Y, & Fikrig E. (2022). A tick C1q protein alters infectivity of the Lyme disease agent by modulating interferon γ. Cell reports, 41(8), 111673.
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2

Tick Bite Immunological Response in Mice

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After 48 h tick bite (n = 10 per mouse), WT (n = 6) and KO mice (n = 6) were euthanized as described above, and one 3.5 mm (Integra LifeSciences, US) punch biopsy specimen was taken from tick bite site on the ears. Total RNA was extracted using RNeasy Fibrous Tissue Mini Kit according to the manufacturer’s instructions (Qiagen, # 74704). cDNA was synthesized, and qPCR was performed as described above. In addition, we also quantify cytokines production by the Mouse Cytokine/Chemokine Array 32-plex (MD-32) performed by Eve Technologies. Serum collected from each group of mice was sent for cytokine analyses. The cytokines represented by this array are Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, KC, LIF, LIX, MCP-1, M-CSF, MIG, MIP-1α, MIP-1β, MIP-2, RANTES, and TNFα.
Tang X., Cao Y., Booth C.J., Arora G., Cui Y., Matias J, & Fikrig E. (2023). Adiponectin in the mammalian host influences ticks’ acquisition of the Lyme disease pathogen Borrelia. PLOS Biology, 21(10), e3002331.
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3

Investigating the Immunomodulatory Effects of IsC1ql3

Check if the same lab product or an alternative is used in the 5 most similar protocols
We determined the effects of IsC1ql3 on host cytokine production in vitro and in vivo. For in vitro assays, the murine splenocytes, CD4+ and CD8+ T cells were isolated as described above. The isolated cells were incubated with 1 μg/mL rIsC1ql3 or BSA (control) and stimulated by 10 ng/mL LPS or 106B. burgdorferi for 6 h. The cells and supernatant were harvested, and total RNA was extracted using a RNeasy Mini kit (Qiagen). For in vivo studies, tick IsC1ql3 gene was silenced as described above. Then the silenced nymphal ticks were put on mice ears. After 72 h post tick bite, mice were be sacrificed, and punch biopsy specimens were taken from the tick bite site. Total RNA was extracted, and cDNA was synthesized as described above. The qPCR primers sequences of cytokine genes are shown in Table S2. In addition, we also quantify cytokines production by the Mouse Cytokine/Chemokine Array 32-plex (MD-32) performed by Eve Technologies. Serum collected from each group of mice was sent for cytokine analyses. The cytokines represented by this array are Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, KC, LIF, LIX, MCP-1, M-CSF, MIG, MIP-1α, MIP-1β, MIP-2, RANTES, TNFα, and VEGF.
Tang X., Arora G., Matias J., Hart T., Cui Y, & Fikrig E. (2022). A tick C1q protein alters infectivity of the Lyme disease agent by modulating interferon γ. Cell reports, 41(8), 111673.
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4

Measuring Neuroendocrine and Neurochemical Changes in Chronic Stress

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Fasted glycemia was measured in 6-h-fasted mice at the end of the UCMS procedure as previously described [49 (link)]. One week after completion of behavioral testing, mice were anesthetized by isoflurane inhalation and blood samples immediately collected via cardiac puncture [49 (link)]. Commercial kits were used to assay plasma corticosterone (Corticosterone-HS kit; ImmunoDiagnostic System, France), leptin, resistin and adiponectin concentrations (Metabolic- and Adiponectin-Milliplex kits; Merck-Millipore, France). Plasma chemokine and cytokine assays were conducted by Eve Technologies (Calgary, Canada) using a bead-based multiplex assay (Mouse Cytokine/Chemokine Array 32-Plex [MD32]). After blood collection, mice were perfused with chilled 1X PBS and part of the brains rapidly dissected to collect and immediately frozen the hippocampus (HC), prefrontal cortex (PFC) and striatum. After homogenization, DA, 5-HT and their main metabolites (dihydroxyphenyl acetic acid (DOPAC), homovanillic acid (HVA), 5-hydroxyindoleacetic acid (5-HIAA)) were measured by HPLC-EC [50 (link)]. Brains used to measure mRNA expression were directly stored at − 80 °C until they were micropunch-dissected as previously described [51 ].
Cardinal P., Monchaux de Oliveira C., Sauvant J., Foury A., Darnaudéry M., Vancassel S., Castanon N, & Capuron L. (2021). A new experimental design to study inflammation-related versus non-inflammation-related depression in mice. Journal of Neuroinflammation, 18, 290.
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5

Tick Bite Immunological Response in Mice

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After 48 h tick bite (n = 10 per mouse), WT (n = 6) and KO mice (n = 6) were euthanized as described above, and one 3.5 mm (Integra LifeSciences, US) punch biopsy specimen was taken from tick bite site on the ears. Total RNA was extracted using RNeasy Fibrous Tissue Mini Kit according to the manufacturer’s instructions (Qiagen, # 74704). cDNA was synthesized, and qPCR was performed as described above. In addition, we also quantify cytokines production by the Mouse Cytokine/Chemokine Array 32-plex (MD-32) performed by Eve Technologies. Serum collected from each group of mice was sent for cytokine analyses. The cytokines represented by this array are Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, KC, LIF, LIX, MCP-1, M-CSF, MIG, MIP-1α, MIP-1β, MIP-2, RANTES, and TNFα.
Tang X., Cao Y., Booth C.J., Arora G., Cui Y., Matias J, & Fikrig E. (2023). Adiponectin in the mammalian host influences ticks’ acquisition of the Lyme disease pathogen Borrelia. PLOS Biology, 21(10), e3002331.
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+ Expand

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