Cxcl2 mip 2

Manufactured by R&D Systems

Sourced in United States

CXCL2/MIP-2 is a chemokine that is involved in the recruitment and activation of neutrophils. It is a member of the CXC chemokine family.

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Lab products found in correlation

Cxcl1 kc, by R&D Systems (4 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Fixation and permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Rabbit anti mouse relmα, by Thermo Fisher Scientific (1 mentions) Cd86 pe, by Thermo Fisher Scientific (1 mentions) Rat igg, by Merck Group (1 mentions) F4 80 percp cy5, by BioLegend (1 mentions) Ly6c apc cy7, by BioLegend (1 mentions) Softmax pro 6, by Molecular Devices (1 mentions) Alexa fluor 647, by BioLegend (1 mentions) Ly6g pe, by BioLegend (1 mentions) Tumor necrosis factor (tnf), by Thermo Fisher Scientific (1 mentions) Tgf β, by Thermo Fisher Scientific (1 mentions) Mhcii apc, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Aposcreen annexin 5 fitc kit, by Southern Biotech (1 mentions) Penicillin, by Merck Group (1 mentions) Rpmi 1640 medium, by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) Lipopolysaccharide, by Merck Group (1 mentions)

6 protocols using cxcl2 mip 2

Flow Cytometric Analysis of Immune Cells

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For flow cytometric analysis, cells were blocked with PBS containing 1% bovine serum albumin and 0.1% rat IgG (Sigma-Aldrich, St. Louis, USA) for 30 min. After a washing step, cells were stained with SiglecF-PE or -AlexaFluor647 (Clone: E50-2440, BD Pharmingen, San Diego, USA), F4/80-PerCP-Cy5.5 (Clone: BM8), Ly6G-PE (Clone: 1A8), Ly6C-APC-Cy7 (Clone HK1.4) (all BioLegend, San Diego, USA), and CD11b-FITC or -PE-Cy7 (Clone: M1/7; eBioscience, San Diego, USA). For intracellular staining, cells were incubated with fixation and permeabilization buffer (eBioscience) overnight. Cells were stained with rabbit anti-mouse RELMα (Peprotech, Rocky Hill, USA) followed by donkey anti-rabbit IgG AlexaFluor647 (Clone: Poly4064, BioLegend) and CD86-PE (Clone: GL1) and MHCII-APC (Clone: M5/114.15.2, eBioscience) to determine cell activation. The gating strategy used to identify macrophages, monocytes, eosinophils and neutrophils is shown in Figure 1 (Fig. 1).
IFNγ , IL-10, TGFβ and TNF (all eBioscience) as well as CXCL1/KC and CXCL2/MIP-2 (both R&D, Minneapolis, USA) were measured from peritoneal lavage and serum by ELISA according to the manufacturers’ protocols and analyzed using a plate reader (Molecular Devices) with SoftMax Pro 6.

Buerfent B.C., Ajendra J., Stamminger W., Gondorf F., Hoerauf A, & Hübner M.P. (2019). TGFβ depletion does neither modulate acute E. coli-induced inflammatory immune responses nor impair the protective effect by chronic filarial infection. GMS Infectious Diseases, 7, Doc04.

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Neovestitol Antiinflammatory Activity Assessment

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Ethanol, hexane, chloroform and ethyl acetate were purchased from Merck (Sao Paulo, SP, Brazil). Lipopolysaccharide (LPS), dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), RPMI-1640 Medium, L-glutamine and penicillin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Type II bovine collagen from MD Biosciences (St. Paul, MN, USA). Apo Screen Annexin V-FITC Kit was purchased from SouthernBiotech (Birmingham, AL, USA). CXCL2/MIP-2 from R&D Systems (Minneapolis, MN, USA). Fetal bovine serum from Gibco (Grand Island, NY, USA). Neovestitol was dissolved in 1% DMSO in PBS. The negative control group received treatment with vehicle alone (1% DMSO in PBS).

Franchin M., Colón D.F., da Cunha M.G., Castanheira F.V., Saraiva A.L., Bueno-Silva B., Alencar S.M., Cunha T.M, & Rosalen P.L. (2016). Neovestitol, an isoflavonoid isolated from Brazilian red propolis, reduces acute and chronic inflammation: involvement of nitric oxide and IL-6. Scientific Reports, 6, 36401.

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Quantification of Inflammatory Cytokines

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Mouse IL-1β (BD Biosciences, North Ryde, NSW, Australia), IL-18 (R&D Systems, In Vitro Technologies Pty Ltd., VIC, Australia), Cxcl1/KC (R&D Systems), Cxcl2/MIP2 (R&D Systems), human IL-18 (R&D Systems) and CXCL8 (BD Biosciences) were quantified by ELISA, as per the manufacturers’ instructions. Production of bioactive IL-18 by AGS cells was also measured using the HEK-Blue™ IL-18 reporter cell line system (InvivoGen, San Diego, CA, USA).

Tran L.S., Ying L., D’Costa K., Wray-McCann G., Kerr G., Le L., Allison C.C., Ferrand J., Chaudhry H., Emery J., De Paoli A., Colon N., Creed S., Kaparakis-Liaskos M., Como J., Dowling J.K., Johanesen P.A., Kufer T.A., Pedersen J.S., Mansell A., Philpott D.J., Elgass K.D., Abud H.E., Nachbur U., Croker B.A., Masters S.L, & Ferrero R.L. (2023). NOD1 mediates interleukin-18 processing in epithelial cells responding to Helicobacter pylori infection in mice. Nature Communications, 14, 3804.

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Intranasal Pneumococcus Infection in Mice

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Age-matched 8–12 week old female MF1 mice (Charles River, UK) were intranasally infected with 1 × 10⁵ colony forming units of S. pneumoniae in 10 μl PBS as previously described [28] (link). Mice were sacrificed at pre-determined time points post-infection and organs removed for assessment of bacterial numbers and ELISA analysis. Nasopharynx and lungs were homogenized in PBS and serially diluted onto blood agar for enumeration of bacterial numbers by the Miles and Misra method. For the CFU counting, gentamicin plates were used to select for pneumococci and in the case of the bacteria that disseminated to the lung, colonies were picked and streaked onto plates with an optochin disc to confirm that they were pneumococci. Homogenates were retained for use in ELISA to measure a murine homologue of human CXCL8, CXCL2/MIP-2, using a kit from R&D systems.

Küng E., Coward W.R., Neill D.R., Malak H.A., Mühlemann K., Kadioglu A., Hilty M, & Hathaway L.J. (2014). The Pneumococcal Polysaccharide Capsule and Pneumolysin Differentially Affect CXCL8 and IL-6 Release from Cells of the Upper and Lower Respiratory Tract. PLoS ONE, 9(3), e92355.

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Cell Viability Assay and Immunological Evaluation

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Alamar Blue cell viability reagent was purchased from Invitrogen (Massachusetts, USA). Dexamethasone, diclofenac sodium, dimethyl sulfoxide (DMSO), Dulbecco's Modified Eagle Medium (DMEM), ethylenediaminetetraacetic acid (EDTA), fetal bovine serum (FBS), Laemmli sample buffer, lipopolysaccharide (LPS), phosphate-buffered saline (PBS), and protease inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ether, ethanol, ethyl acetate, dichloromethane, methanol, n-butanol, and n-hexane were purchased from Tedia Way (Fairfield, OH, USA). Nitrocellulose membranes were purchased from Amersham Pharmacia Biotech (San Francisco, CA, USA). The chromatographic solvents (acetonitrile, acetic acid, and trifluoroacetic acid) were purchased from Merck (Darmstadt, Germany). Monoclonal antibodies anti-COX-2, anti-goat IgG biotin-conjugated antibody, and streptavidin-conjugated horseradish peroxidase were purchased from Santa Cruz Biotechnology (Texas, USA). The CXCL2/MIP-2, CXCL1/KC, CCL11/eotaxin-1, and TNF-α enzyme-linked immunosorbent assay (ELISA) kit were purchased from R&D Systems (Minneapolis, MN, USA). The colorimetric kit used for the May-Grunwald-Giemsa method was purchased from Laborclin (Pinhais, PR, BR).

de Brito T.M., Amendoeira F.C., de Oliveira T.B., Doro L.H., Garcia E.B., da Silva N.M., da Silva Chaves A., Muylaert F.F., Pádua T.A., Rosas E.C., Henriques M.D., da Silva Frutuoso V., Valverde S.S, & Ferraris F.K. (2021). Anti-Inflammatory Activity and Chemical Analysis of Different Fractions from Solidago chilensis Inflorescence. Oxidative Medicine and Cellular Longevity, 2021, 7612380.

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Inhibition of IL-1R1/MyD88 Signaling

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The peptide α₂PI_1-8-MyD88-I (NQEQVSPLVPMSMRGGRQIKIWFQNRRMKWKKRDVLPGTCVNS) was synthesized by GeneScript and verified as ≥95.0% by HPLC. The peptide was further dialyzed against HEPES buffer (20 mM HEPES, 150 mM NaCl, pH 7.5). The capacity of the peptide to inhibit IL-1R1/MyD88 signalling was assessed by monitoring the release of cytokines from MSCs following stimulation with murine IL-1β. MSCs seeded in 24-well at 70–80% confluency were stimulated with 10 ng ml⁻¹ of IL-1β together with α₂PI_1-8-MyD88-I at increasing concentration (1–100 μg ml⁻¹). After 24 h, the concentration of IL-6, CCL2, CXCL1 and CXCL2 released were measured using ELISA (Mouse IL-6, CCL2/JE/MCP-1, CXCL1/KC and CXCL2/MIP-2; DuoSets, R&D Systems).

Martino M.M., Maruyama K., Kuhn G.A., Satoh T., Takeuchi O., Müller R, & Akira S. (2016). Inhibition of IL-1R1/MyD88 signalling promotes mesenchymal stem cell-driven tissue regeneration. Nature Communications, 7, 11051.

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