Met rantes

Manufactured by R&D Systems

Sourced in United States

Met-RANTES is a recombinant human chemokine. It is a chemotactic and activating factor for a variety of cell types, including monocytes, eosinophils, and T cells.

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Lab products found in correlation

Rantes, by Thermo Fisher Scientific (1 mentions) α galcer, by Enzo Life Sciences (1 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Cell Signaling Technology (1 mentions) Mouse ifn γ elisa kit, by BioLegend (1 mentions) Murine il 3, by Thermo Fisher Scientific (1 mentions) Murine il 6, by Thermo Fisher Scientific (1 mentions) Mouse il 4 elisa kit, by BioLegend (1 mentions) Collagenase 4, by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Taq master mix, by Vazyme (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Heparinase 3, by Merck Group (1 mentions) Total p38, by Cell Signaling Technology (1 mentions) Mmp 13, by Santa Cruz Biotechnology (1 mentions) Mmp 1, by Santa Cruz Biotechnology (1 mentions) Mmp 1 elisa duoset, by R&D Systems (1 mentions) P c jun ser73, by Cell Signaling Technology (1 mentions) Total jnk, by Cell Signaling Technology (1 mentions) Collagen type 1, by Santa Cruz Biotechnology (1 mentions)

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RANTES

5 protocols using met rantes

CCL5 and Ezrin Inhibition in Cancer

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For CCL5 inhibition, met-RANTES (R&D systems, MN, USA), a recombinant protein, which is shown to be a partial agonist in the monocyte chemotactic assays, was used. The cells were treated with 100 ng metCCL5 and the inhibition of CCL5 was confirmed by qRT-PCR. Cancer cells or CAFs cultured in presence of cmCAFs or cmHCC1937 and cmHCC1937/wt BRCA1 respectively were treated with or without metCCL5 for 48 h and then used for further analysis.
For Ezrin inhibition, pBS/U6 ezrin siRNA, which was a gift from Philip Hinds (Addgene plasmid # 8945) were used for transiently transfecting cancer cells/CAFs using lipofectamine reagent. The inhibition was confirmed with qRT-PCR. Further ELISA, migration and invasion assays were done.

Hemalatha S.K., Sengodan S.K., Nadhan R., Dev J., Sushama R.R., Somasundaram V., Thankappan R., Rajan A., Latha N.R., Varghese G.R., Mathew A.P., Somanathan T, & Srinivas P. (2018). Brcal Defective Breast Cancer Cells Induce in vitro Transformation of Cancer Associated Fibroblasts (CAFs) to Metastasis Associated Fibroblasts (MAF). Scientific Reports, 8, 13903.

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Retrograde Perfusion of Rat Vas Deferens

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Adult male Sprague-Dawley rats were anesthetized with sodium pentobarbital as described above. The vas deferens was cannulated through the lumen with a micro cannula (0.31 mm OD, 0.16 mm ID; Anilab Software & Instruments Co., PE-0402) connected to a 1-ml syringe. A small incision was made in the distal cauda epididymal region to allow the perfusate to exit the tubule. Perfusion was performed retrogradely at a rate of 20 μl/min using a syringe pump (78-0120S, Stoelting). The lumen was initially washed free of sperm with PBS (0.01M, pH 6.8) for usually 10 min. Then, vas deferens and cauda epididymis were perfused with recombinant RANTES (PeproTech, United States), Met-RANTES (R&D Systems, United States), and PBS of different pH values, respectively. The perfusion was performed retrogradely at a rate of 5 μl/min, and the luminal solution was sustained for 1 h. At the end of the experimental period, the cauda epididymis was harvested and extracted for the RNA and the protein. Or tissues were then washed in PBS, pH 7.4, and perfused with a solution containing 4% paraformaldehyde for 15 min for frozen sections.

Feng X., Ma B.F., Liu B., Ding P., Wei J.H., Cheng P., Li S.Y., Chen D.X., Sun Z.J, & Li Z. (2020). The Involvement of the Chemokine RANTES in Regulating Luminal Acidification in Rat Epididymis. Frontiers in Immunology, 11, 583274.

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Multiparameter Flow Cytometry Analysis

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Fluorescently conjugated protein or antibodies used for cell-surface staining and intracellular staining for cytokines and transcription factors are listed in Supplementary Table 2. α-GalCer (KRN7000) was purchased from Enzo. CD1d-PBS57 (conjugated with either PE or allophycocyanin) was obtained from the tetramer facility of the US National Institutes of Health. Collagenase IV used for tissue digestion was purchased from Sigma. Taq Master Mix was purchased from Vazyme Biotech. A mouse IFN-γ ELISA kit and a mouse IL-4 ELISA kit were obtained from BioLegend. PMA and ionomycin were obtained from Merck. Stem cell factor, murine IL-3, and murine IL-6 were obtained from PeproTech. Met-RANTES was purchased from R&D Systems. DAPI was obtained from Cell Signaling Technology.

Xu Y., Sun Y., Shen H., Dai Y., Liu H., Li R., Zhang H., Wu L., Zhu X, & Liu X. (2018). Regulation of the terminal maturation of iNKT cells by mediator complex subunit 23. Nature Communications, 9, 3875.

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RANTES and MMP Regulation in Fibrosis

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Recombinant human RANTES/CCL5, Met-RANTES and IL-1β, and MMP-1 ELISA Duoset were purchased from R&D systems (Minneapolis, MN, USA). MMP-13 ELISA was purchased from Ray Biotech (Norcross, GA, USA). MMP-1 and MMP-13 SYBR Green primers were purchased from Qaigen (Valencia, CA, USA). SMARTpool ON-TARGETplus RANTES small-interfering RNA (siRNA) and On-target plus non targeting siRNA (D-001810-10) control were purchased from GE Dharmacon (Lafayette, CO, USA). MMP-1, MMP-13, type I collagen, and lamin A/C antibodies were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA). Rabbit monoclonal or polyclonal antibodies against p-p38 (Thr¹⁸⁰/Tyr¹⁸²), p-ERK (Thr²⁰²/Tyr²⁰⁴), p-JNK(Thr¹⁸³/Tyr¹⁸⁵), p-PKCδ (Thr⁵⁰⁵), total p-38, total JNK, total ERK, p-c-Jun (Ser⁷³), and p-ATF-2 (Thr^69/71) were from Cell Signaling Technologies (Beverly, MA, USA). Rabbit polyclonal antibodies against CCR1 and CCR5 were from BioVision (Milpitas, CA) and GeneTex (Irvine, CA, USA), respectively. Heparinase III (flavobacterium heparinum derived), heparan sulfate degrading lyase was purchased from Sigma (St. Louis, MO, USA). type I collagen was purchased from Advanced Biometrix (Carlsbad, CA, USA). Inhibitors of JNK (SP600125), ERK (PD98059), p38 (SB 02190), PKCδ (Rottlerin), and NF-κB (PDTC) were purchased from EMD Millipore (Billerica, MA, USA).

Agere S.A., Akhtar N., Watson J.M, & Ahmed S. (2017). RANTES/CCL5 Induces Collagen Degradation by Activating MMP-1 and MMP-13 Expression in Human Rheumatoid Arthritis Synovial Fibroblasts. Frontiers in Immunology, 8, 1341.

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Lung Cancer Cell Sensitization to Chemotherapy

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NSCLC A549 (lung adenocarcinoma) and H1299 (lung large cell carcinoma) cell lines were purchased from the China Center for Type Culture Collection, and cultivated in RPMI-1640 medium supplemented with 10% FBS, 100 U/ml penicillin and 100 µg/ml streptomycin in a humidified incubator with 5% CO₂ at 37°C. Cells in the logarithmic growth phase were used for all experiments.
For cell treatment, cancer cells were incubated with CAF-CM or NF-CM in combination with either anti-CCL5 antibody (0.1 µg/ml; cat. no. MAB678-SP; R&D Systems, Inc.), CCR5 antagonist (Met-RANTES; 0.1 µg/ml; cat. no. 335-RM-025; R&D Systems, Inc.) or recombinant human CCL5 (3 ng/ml; cat. no. 300-06; PeproTech, Inc.) for 6 h, followed by treatment with 50 µM DDP (Sigma-Aldrich; Merck KGaA) in the presence of CM for another 48 h.

Sun X, & Chen Z. (2021). Cancer-associated fibroblast-derived CCL5 contributes to cisplatin resistance in A549 NSCLC cells partially through upregulation of lncRNA HOTAIR expression. Oncology Letters, 22(4), 696.

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