ZA was sourced from Novartis Pharma AG Basel, Switzerland. Monoclonal antibodies against human CD25-PE were purchased from eBioscience (San Diego, CA, USA), CD69-FITC was sourced from BD Biosciences (San Jose, CA, USA); the secondary antibody goat anti-mouse IgG FITC was sourced from Merck Millipore (Billerica, MA); and human CCL2/MCP1, human CCL5/RANTES, anti-CCL2 antibody, and anti-CCL5 antibody were sourced from R & D Systems (Minneapolis MN). The triple negative breast cancer cell lines MDA-MB-231 and MDA-MB-468 were purchased from Food Industry Research and Development Institute (FIRDI) and Culture Collection and Research Center (CCRC, Taiwan, ROC). Both cell line were cultured with DMEM medium (Gibco®, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; Biological Industries, Kibbutz Beit HaEmek, Israel).
Human ccl2 mcp 1
Manufactured by R&D Systems
Sourced in United States
Human CCL2/MCP-1 is a recombinant human chemokine protein. It is a chemoattractant for monocytes and basophils and plays a role in the inflammatory response.
Automatically generated - may contain errors
Lab products found in correlation
Human ccl5 rantes, by R&D Systems (1 mentions)
Anti ccl2 antibody, by R&D Systems (1 mentions)
Anti ccl5 antibody, by R&D Systems (1 mentions)
Goat anti mouse igg fitc, by Merck Group (1 mentions)
Cd69 fitc, by BD (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Human cxcl8 il 8, by R&D Systems (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Rpmi medium, by Thermo Fisher Scientific (1 mentions)
Heparin, by Merck Group (1 mentions)
Lipopolysaccharide from escherichia coli o111 b4, by Merck Group (1 mentions)
4 protocols using human ccl2 mcp 1
1
Detecting Triple Negative Breast Cancer Markers
2
Quantifying Cytokine Secretion in Cells
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Levels of secreted MCP-1 and IL-8 were measured by determining their concentration in conditioned medium using a commercially available sandwich enzyme-linked immunosorbent assay (ELISA, Human CCL2/MCP-1 and Human IL-8/CXCL8 Immunoassay, R&D systems, Minneapolis, MN, USA), as previously described38 (link). Conditioned medium was harvested on 12 h and 2 days PI, cleared by centrifugation, and stored at − 70 °C. Conditioned medium was acid-activated and directly assayed using an ELISA plate reader at 450 nm, according to the manufacturer’s instructions. Protein concentrations were calculated from a standard curve with two-fold serial dilutions and a highest standard of 2000 pg/ml.
3
Cytokine Release from Platelet-Derived EVs
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Cytokine release in response to isolated platelet-derived EVs was measured in whole blood using commercial ELISA kits. Whole blood from five healthy donors was diluted 1:5 in RPMI medium (Gibco, Life Technologies) and stimulated with 50 μL isolated EVs from platelets stimulated with HEPES, thrombin, or M1 protein for 24 h in a CO2 incubator (37°C, 5% CO2, >95% relative humidity). Heparin (0.6 μg/mL, Sigma-Aldrich) was added in combination with penicillin (100 mg/mL, Gibco, Life Technologies) and streptomycin (100 mg/mL, Gibco, Life Technologies) to prevent clot formation and contamination. Lipopolysaccharide from Escherichia coli O111:B4 (1 μg/mL, EMD Millipore Corp.) was used as a positive control for monocyte activation, and HEPES buffer alone was used to determine the background cytokine release. The cytokine release mediated by M1 protein alone (2.5 μg/mL) was also investigated. After incubation, the samples were centrifuged at 500 g for 5 min, and cytokine and chemokine levels in the supernatants were measured using commercial ELISA kits for IL-6 (Invitrogen, Thermo Fisher Scientific), human CCL2/MCP-1 (R and D Systems), and human CXCL8/IL-8 (R and D Systems), according to manufacturer instructions.
4
Ovarian Function and Systemic Inflammation
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