Mcp 1

Manufactured by RayBiotech

Sourced in United States

MCP-1 is a protein that functions as a chemotactic factor, responsible for the recruitment and activation of monocytes and other immune cells to sites of inflammation or injury. It is a key mediator of the inflammatory response.

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17 protocols using mcp 1

Quantifying MCP-1 and CXCL5 Levels

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Cell culture supernatant samples were centrifuged for 10 min (6000 r/min, 4°C). MCP-1 (product number: ELR-MCP1, RayBiotech, Norcross, GA, USA) commercial sandwich ELISA Kit was used to test MCP-1 protein levels. CXCL5 (product number: KA1828, Abnova, Taiwan) commercial sandwich ELISA Kit was used to measure CXCL5 protein levels.

Wu C., Qin N., Ren H., Yang M., Liu S, & Wang Q. (2018). Metformin Regulating miR-34a Pathway to Inhibit Egr1 in Rat Mesangial Cells Cultured with High Glucose. International Journal of Endocrinology, 2018, 6462793.

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Cytokine Secretion Profiling in RAOEC

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RAOEC cells were cultured in a 6-well plate and treated with supplemented DMEM in the following groups: C, T, SF/CS, ETS/CSL, ETS/CSH, and CS (the intervention concentration of each group was the same as that of cell migration experiment). After 24 h, the cell culture supernatant was extracted, and enzyme-linked immunosorbent assay (ELISA) kits for detecting monocyte chemoattractant protein-1 (MCP-1) (SAB, USA), granulocyte-macrophage colony-stimulating factor (GM-CS) (Fcmacs, Nanjing), interleukin-8 (IL-8) (Fcmacs), and IP10 (RayBio, USA) were used to detect MCP-1, GM-CSF, IL-8, and IP10 levels, respectively, according to their respective manufacturer’s instructions.

Tu P., Pan Y., Wang L., Li B., Sun X., Liang Z., Liu M., Zhao Z., Wu C., Wang J., Wang Z., Song Y., Zhang Y., Ma Y, & Guo Y. (2024). CD62E- and ROS-Responsive ETS Improves Cartilage Repair by Inhibiting Endothelial Cell Activation through OPA1-Mediated Mitochondrial Homeostasis. Biomaterials Research, 28, 0006.

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Quantification of Ginsenosides and Monosaccharides

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The reference substances of ginsenosides (nR1, Rg1, Re, pF11, Rf, Ra2, Rb1, Rc, Ro, Rb2, Rb3, Rd and 20(R)-Rg3) and monosaccharides (Glc, Ara, Gal, Man, GalA, Rha, Rib, ClcUA, Xyl and Fuc) were purchased from Shanghai Yuanye Biotechnology (Shanghai, China). The purity of these references was higher than 99.0% indicated by HPLC analysis. Cyclophosphamide (CP) was supplied by Sigma-Aldrich Company Ltd., (United States). Thrombopoietin (TPO), erythropoietin (EPO), and granulocyte-macrophage colony stimulating factor (GM-CSF) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Quantibody®Array Glass Chip of IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-13, IFNγ, TNF-α and MCP-1 were purchased from RayBiotech Life, Inc. (United States). FITC-Annexin-V and Cell Cycle kits were purchased from BD Biosciences Pharmingen (United States). The blood cell analysis reagent kit was purchased from IDEXX Laboratories Inc (United States). Protopanaxadiol saponins (PDS) and protopanaxatriol saponins (PTS) were supplied by Professor Chen Yanping in the College of Chemistry, Jilin University; PDS and PTS were LC-MS grade. The voucher specimen of PDS (S20190011) and PTS (S20190012) were deposited in the authors’ lab in Changchun University of Chinese Medicine (Changchun, China).

Zhang H., Zhang L., Yang C., Zhang Y., Li J., Zhang X., Chen J., Huang B., Zhao D., Li X., Zhang W, & Qi B. (2022). Prevention Effect of Protopanaxadiol-Type Saponins Saponins and Protopanaxatriol-Type Saponins on Myelosuppression Mice Induced by Cyclophosphamide. Frontiers in Pharmacology, 13, 845034.

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Cytokine Quantification in Cell Supernatants

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Cell culture supernatants were collected after each treatment, centrifuged and stored at −80 °C until use for MCP-1 (RayBiotech Life, Inc., Peachtree Corners, GA, USA) and IL-8 (Invitrogen, Waltham, MA, USA) quantification by ELISA, according to the manufacturer’s protocol.

Giordani C., Matacchione G., Giuliani A., Valli D., Scarpa E.S., Antonelli A., Sabbatinelli J., Giacchetti G., Sabatelli S., Olivieri F, & Rippo M.R. (2023). Pro-Osteogenic and Anti-Inflammatory Synergistic Effect of Orthosilicic Acid, Vitamin K2, Curcumin, Polydatin and Quercetin Combination in Young and Senescent Bone Marrow-Derived Mesenchymal Stromal Cells. International Journal of Molecular Sciences, 24(10), 8820.

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Multiplex Analysis of Rat Synovial Inflammation

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Rats were euthanized via CO₂ inhalation at three days or twenty-one days post-surgery for both the naïve group and the MMT surgery group. Synovial fluid was collected by first injecting 100 μl of saline intra-articularly using a 30 gauge insulin syringe, followed by aspirating approximately 50 to 100 μl of the synovial fluid and saline using the same syringe. Synovial fluid was analyzed using the Quantibody^® Rat Inflammation Array 1, a multiplex ELISA kit that quantitatively measured 10 rat inflammatory factors: IFNγ, IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-13, monocyte chemoattractant protein-1 (MCP-1), and TNFα (RayBiotech, Norcross, GA, USA).

Willett N.J., Thote T., Lin A.S., Moran S., Raji Y., Sridaran S., Stevens H.Y, & Guldberg R.E. (2014). Intra-articular injection of micronized dehydrated human amnion/chorion membrane attenuates osteoarthritis development. Arthritis Research & Therapy, 16(1), R47.

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Inflammasome Activation in Immune Cells

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LPS and pancreatic enzymes were purchased from Sigma-Aldrich (St. Louis, MO, USA); lymphocyte separation medium was purchased from Jingyang Biological (Tianjin, China); fetal bovine serum (FBS) was obtained from GIBCO (Australia); RPMI-1640 glucose free medium was purchased from Hyclone (Logan, UT, USA); RNeasy Mini Kit, RNase-Free DNase Set, RT² First Strand Kit, RT² SYBR® Green ROX qPCR Mastermix, and Human Inflammasomes PCR Array were obtained from Qiagen (Valencia, CA, USA). ELISA kits of human TNF-α, IL-1β, IL-6, IL-10, and MCP-1 were purchased from RayBiotech (Atlanta, USA). Anti-NLRP3 and anti-Caspase-1 antibodies were purchased from Abcam (Cambridge, United Kingdom). Secondary antibodies were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Sai J., Xiong L., Zheng J., Liu C., Lu Y., Wang G., Wang Y., Wang T., Guan X., Chen F., Fang K., Zhang C., Lu J., Zhang X., Zhu H, & Wang F. (2016). Protective Effect of Yinhua Miyanling Tablet on Lipopolysaccharide-Induced Inflammation through Suppression of NLRP3/Caspase-1 Inflammasome in Human Peripheral Blood Mononuclear Cells. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 2758140.

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Quantification of Inflammatory Markers

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Circulating and tissue levels of Activin A (R&D Systems), β‐2 microglobulin (β2M, Abcam), GDF15 (R&D Systems), IL‐1β (Abcam), MCP‐1 (RayBiotech), and Osteopontin (OPN, R&D Systems) were measured by ELISA using a Varioskan plate reader (Thermo Fisher) according to manufacturer's specifications.

Yousefzadeh M.J., Zhao J., Bukata C., Wade E.A., McGowan S.J., Angelini L.A., Bank M.P., Gurkar A.U., McGuckian C.A., Calubag M.F., Kato J.I., Burd C.E., Robbins P.D, & Niedernhofer L.J. (2020). Tissue specificity of senescent cell accumulation during physiologic and accelerated aging of mice. Aging Cell, 19(3), e13094.

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Profiling Inflammatory Biomarkers in Mice

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Blood from mice, as well as medium supernatant were collected for measuring the concentrations of interleukin- (IL-) 8, monocyte chemotactic protein-1 (MCP-1), IL-1α, IL-6, vascular endothelial growth factor (VEGF) (RayBiotech, GA, US), transforming growth factor-beta (TGF-β), C-reactive protein (CRP), MMP-3, MMP-9 (R&D Systems, MN, US) according to manufacturer’s instructions.

Lou Z., Zhu J., Li X., Li X., Du K., Wang B., Zhang F, & Zhang X. (2021). LncRNA Sirt1-AS upregulates Sirt1 to attenuate aging related deep venous thrombosis. Aging (Albany NY), 13(5), 6918-6935.

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Quantifying Inflammatory and Anti-Inflammatory Cytokines

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Pro-inflammatory cytokines (RayBiotech, MCP-1 #ELM-MCP-001 and TNF-α #ELM-TNF-α-001), anti-inflammatory cytokines (RayBiotech, IL-10 #ELM-IL10-001 and IL-1RA #ELM-IL1RA-001), and BMP-7 expression (TSZ Scientific, #M7485) were quantified using commercially available ELISA kits, per manufacture’s instructions, on blood serum samples previously collected from each animal. In brief, blood from each animal was collected post-sacrifice in EDTA coated tubes and centrifuged at 13000 rpm for 15 min. Supernatant containing serum was collected into separate tubes and stored at -20°C for future analysis. For each ELISA, serum was loaded into pre-coated 8-well strips for 2.5 hrs at room temperature. After washing, the detection antibody (biotinylated anti-mouse antibodies) was added followed by HRP-conjugated streptavidin and TMB One-Step substrate reagent, as provided in the kit. When the blue color developed, stop solution was added and the wells were measured at 450 nm in a microtiter plate reader (Bio Rad). Results were normalized to protein concentration and data is presented as arbitrary units (A.U.).

Singla D.K., Singla R, & Wang J. (2016). BMP-7 Treatment Increases M2 Macrophage Differentiation and Reduces Inflammation and Plaque Formation in Apo E-/- Mice. PLoS ONE, 11(1), e0147897.

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Cytokine Profiling of Injured Rat Aortas

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Injured rat aortas or HMAs were recovered and homogenized in cell lysing buffer (including proteinase inhibitor) using the Qiagen Tissue Lyser. Tissue lysate (400 µg) was diluted in 1 ml assay buffer and membranes were incubated for 2 h at 20–25 °C followed by antibody incubation according to the protocol. Cytokine antibody arrays for PDGF, IFNγ, MCP-1, MIP-3α, and IL-1β (Raybiotech, Norcross, Georgia, USA) were performed according to the manufacturer’s protocol. The membranes were digitized using bioluminescence imaging and quantified using NIH ImageJ. Cytokine concentration is expressed in arbitrary units (AU).

Deuse T., Hua X., Wang D., Maegdefessel L., Heeren J., Scheja L., Bolaños J.P., Rakovic A., Spin J.M., Stubbendorff M., Ikeno F., Länger F., Zeller T., Schulte-Uentrop L., Stoehr A., Itagaki R., Haddad F., Eschenhagen T., Blankenberg S., Kiefmann R., Reichenspurner H., Velden J., Klein C., Yeung A., Robbins R.C., Tsao P.S, & Schrepfer S. (2014). Dichloroacetate prevents restenosis in preclinical animal models of vessel injury. Nature, 509(7502), 641-644.

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