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Cytokine bead array mouse inflammation kit

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The Cytokine Bead Array Mouse Inflammation kit is a lab equipment product designed for the simultaneous quantitative measurement of multiple mouse inflammatory cytokines in a single sample. The core function of this kit is to enable the detection and analysis of cytokine levels in various biological samples.

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Lab products found in correlation

Prism 4, by GraphPad (1 mentions) Moloney murine leukaemia virus reverse transcriptase, by Promega (1 mentions) Random hexamer, by Thermo Fisher Scientific (1 mentions) Rnaeasy isolation kit, by Qiagen (1 mentions) Sybr reagents, by Thermo Fisher Scientific (1 mentions) Il 1β, by R&D Systems (1 mentions) Legendplex multiplex assay kit, by BioLegend (1 mentions) Griess reagent, by Promega (1 mentions) Rmm csf, by Thermo Fisher Scientific (1 mentions) Rbc lysis solution, by Thermo Fisher Scientific (1 mentions) Rmil 4, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Cytokine Bead Array (59 citations) Mouse Inflammation Kit (52 citations)

4 protocols using cytokine bead array mouse inflammation kit

1

Cytokine Bead Array Analysis of BMDM

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The Cytokine Bead Array Mouse Inflammation kit (BD Biosciences 552364) was used according to manufacturer’s instructions for simultaneous measurement of IL-6, IL-10, CCL2, IFNγ, TNFα, and IL-12p70 in supernatants from stimulated BMDMs. General statistical analysis was performed using GraphPad Prism 4.0 (GraphPad Software, San Diego, CA), using an unpaired Student t test. For each experiment, both the standard deviation and the standard error of the mean were calculated. P-values of < 0.05 were considered significant.
Benjamin S.J., Hawley K.L., Vera-Licona P., La Vake C.J., Cervantes J.L., Ruan Y., Radolf J.D, & Salazar J.C. (2021). Macrophage mediated recognition and clearance of Borrelia burgdorferi elicits MyD88-dependent and -independent phagosomal signals that contribute to phagocytosis and inflammation. BMC Immunology, 22, 32.
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2

Quantification of Cytokine Expression in Macrophages

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Total RNA from WT and STAT3 SA peritoneal macrophages was isolated using the RNAeasy Isolation Kit (Qiagen, Cat# 74104) and reverse transcribed with random hexamers (Thermo Fisher Scientific, Cat# N8080127) using Moloney murine leukaemia virus reverse transcriptase (Promega, Cat# 1701) according to manufacturers’ instructions. mRNA was quantified with SYBR reagents (Thermo Fisher Scientific, Cat# A46012) using primer pairs targeting il1b (Forward 5′- CAACCAACAAGTGATATTCTCCATG- 3, Reverse 5′- GATCCACACTCTCCAGCTGCA- 3′), and 18S (Forward 5′- GTAACCCGTTGAACCCATT- 3′, Reverse 5′- CGAATCGAATCGGTAGTAGCG- 3′). Relative mRNA expression was analysed using the comparative CT method, normalising genes of interest to the 18S housekeeping gene and fold gene induction calculated relative to expression in control samples.
Quantification of cytokines secreted from peritoneal macrophages and serum, and peritoneal macrophages cellular lysates were conducted using ELISA kits from R&D Systems (IL-1β: Cat# DY401, TNF: Cat# DY410, IL-6: Cat# DY406 and IL-10: Cat# DY417) or Cytokine Bead Array Mouse Inflammation kit (BD Biosciences, Cat# 552364) according to manufacturers’ instructions.
Balic J.J., Albargy H., Luu K., Kirby F.J., Jayasekara W.S., Mansell F., Garama D.J., De Nardo D., Baschuk N., Louis C., Humphries F., Fitzgerald K., Latz E., Gough D.J, & Mansell A. (2020). STAT3 serine phosphorylation is required for TLR4 metabolic reprogramming and IL-1β expression. Nature Communications, 11, 3816.
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3

Cytokine and Nitric Oxide Production in MRSA-Infected Macrophages

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The protein levels of IL-12p70, TNF-α, IFN-γ, MCP-1 and IL-6 were determined using the Cytokine Bead Array Mouse Inflammation Kit (BD Bioscience) and LEGENDplex Multiplex Assay kit (BioLegend, San Diego, CA, USA) following the manufacturer's instructions. IL-10 levels were measured by the Cytokine Beads Array Mouse IL-10 Enhanced Sensitivity Flex Set (BD Bioscience). The NO assay was performed as described (29 (link)). Briefly, macrophages were treated with 5 MOI of MRSA for 8 h, and the culture medium was collected to measure the amount of nitrite, a stable metabolite of NO. One hundred microliters of the culture medium were incubated with equal amount of Griess reagent (Promega, Madison, WI, USA) at room temperature for 10 min, and the absorbance was measured at 540 nm using a microplate reader. The quantity of nitrite was determined from a standard curve of sodium nitrate.
Sim H., Jeong D., Kim H.I., Pak S., Thapa B., Kwon H.J, & Lee K. (2021). CD11b Deficiency Exacerbates Methicillin-Resistant Staphylococcus aureus-Induced Sepsis by Upregulating Inflammatory Responses of Macrophages. Immune Network, 21(2), e13.
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4

Murine Bone Marrow-Derived Macrophage Activation

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To obtain murine BMDMs, femurs and tibias were harvested from female C57Bl/6J mice and marrow cores were flushed using syringes filled with RPMI 1640/10% FBS. Cells were triturated and red blood cells (RBCs) were lysed with 1X RBC lysis solution (eBioscience). Cells were washed once in media, then plated and cultured in RPMI 1640 supplemented with 0.1 mg/mL penicillin/streptomycin, 10 mM HEPES, 0.001% β-mercaptoethanol, 1 mM sodium pyruvate, 2 mM L-glutamine, 0.1 mM non-essential amino acids, and 10% FBS and in the presence of 30 ng/ml rmM-CSF (PeproTech). Cells were incubated for 7 days prior to use. On day 7, cells were scraped and washed with PBS and placed in RPMI/10% FBS. 96-well plates were seeded with 105 BMDMs/well and incubated with 100 μM IA, 100 μM IPA, 100 μM HPPA, or 0.1% DMSO for 6 h, then 20 ng/mL rmIL-4 (PeproTech) was added for 24 h, followed by 20 ng/mL LPS stimulation for 18 h. Supernatants were collected and analyzed using a cytokine bead array mouse inflammation kit (BD Biosciences) according to the manufacturer’s instructions.
Wlodarska M., Luo C., Kolde R., d’Hennezel E., Annand J.W., Heim C.E., Krastel P., Schmitt E.K., Omar A.S., Creasey E.A., Garner A.L., Mohammadi S., O’Connell D.J., Abubucker S., Arthur T.D., Franzosa E.A., Huttenhower C., Murphy L.O., Haiser H.J., Vlamakis H., Porter J.A, & Xavier R.J. (2017). Indoleacrylic acid produced by commensal Peptostreptococcus species suppresses inflammation. Cell host & microbe, 22(1), 25-37.e6.
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