Recombinant human ccl5

Manufactured by R&D Systems

Sourced in United States

Recombinant human CCL5 is a protein produced in a laboratory setting. It is a chemokine that plays a role in the immune system.

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Lab products found in correlation

Anti akt, by Cell Signaling Technology (1 mentions) Ku 57788, by Selleck Chemicals (1 mentions) Anti dna pkcs, by Cell Signaling Technology (1 mentions) Anti γh2ax, by Cell Signaling Technology (1 mentions) Anti p akt ser473, by Cell Signaling Technology (1 mentions) Mk 2206, by Selleck Chemicals (1 mentions) Anti gapdh, by Proteintech (1 mentions) Anti snap tag antibody polyclonal, by New England Biolabs (1 mentions) Affinipure donkey anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions) Anti actin, by Merck Group (1 mentions) Alexa fluor 488 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions) Saponin, by Merck Group (1 mentions) Annexin 5 apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Wortmannin, by Merck Group (1 mentions) Pertussis toxin ptx, by R&D Systems (1 mentions) Retinoic acid, by Merck Group (1 mentions) Cxcl12, by R&D Systems (1 mentions) Mab678 sp, by R&D Systems (1 mentions) Met rantes, by R&D Systems (1 mentions) H1299, by National Collection of Authenticated Cell Cultures (1 mentions)

6 protocols using recombinant human ccl5

Immunoblot Analysis of CCL5-Mediated Signaling

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Immunoblot assay was performed as previously described.^{51 (link)} GBM cells were treated with recombinant human CCL5 (10 ng/mL, R&D) together with or without the AKT inhibitor MK-2206 (Selleckchem, 500 nmol/L), with or without the DNA-PKcs inhibitor KU-57788 (Selleckchem, 1 μmol/L). To block the function of CCR5, MVC was added 1 h before pericyte CM stimulation. Antibodies used for immunoblot assays included anti-AKT (Cell Signaling Technology, #4685, 1:1000), anti-p-AKT-Ser473 (Cell Signaling Technology, #4060, 1:1000), anti-γ-H2AX (Cell Signaling Technology, #9718, 1:1000), anti-DNA-PKcs (Cell Signaling Technology, #38168, 1:1000), anti-p-DNA-PKcs-Ser2056 (Cell Signaling Technology, #68716, 1:1000), anti-GAPDH (Proteintech, #60004, 1:10,000).

Zhang X.N., Yang K.D., Chen C., He Z.C., Wang Q.H., Feng H., Lv S.Q., Wang Y., Mao M., Liu Q., Tan Y.Y., Wang W.Y., Li T.R., Che L.R., Qin Z.Y., Wu L.X., Luo M., Luo C.H., Liu Y.Q., Yin W., Wang C., Guo H.T., Li Q.R., Wang B., Chen W., Wang S., Shi Y., Bian X.W, & Ping Y.F. (2021). Pericytes augment glioblastoma cell resistance to temozolomide through CCL5-CCR5 paracrine signaling. Cell Research, 31(10), 1072-1087.

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Immunofluorescence Labeling of Cell Membrane Proteins

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We obtained anti-human CCR5 45531 from R and D Systems (Minneapolis, MN), anti-actin from Sigma Aldrich (St. Louis, MO), anti-Giantin 924302 from Biolegend (San Diego, CA), Alexa Fluor 488 FluoroNanogold goat anti-mouse IgG Fab from Nanoprobes.com (Yaphank, NY), AffiniPure Donkey Anti-Rabbit IgG (H + L) from Jackson ImmunoResearch (West Grove, PA), Alexa Fluor 488 goat Anti-Mouse IgG (H + L) from Invitrogen (Carlsbad, CA), Anti-SNAP-tag Antibody (Polyclonal) from New England Biolabs (Ipswichm, MA). Hybridoma cells expressing anti-Myc 9E10 were obtained from ATCC. Saponin (from quillaja bark), LPS (from Escherichia coli 0111:B4), glutaraldehyde, and fumonisin B1 (from Fusarium moniliforme) was purchased from Sigma Aldrich (St. Louis, MO). Recombinant human CCL5 was obtained from R and D Systems (Minneapolis, MN).

Denard B., Han S., Kim J., Ross E.M, & Ye J. (2019). Regulating G protein-coupled receptors by topological inversion. eLife, 8, e40234.

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Pericytes Protect GBM Cells from TMZ

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Pericytes (5 × 10⁴) and GBM cells (5 × 10⁴) were, respectively, cultured in the upper or lower chamber in 6-well transwell apparatus, followed by the treatment of TMZ (500 μmol/L, Selleckchem, S1237). To determine the protective effect of pericytes or CCL5 on GBM cells, GBM cells were pretreated with or without pericyte CM, recombinant human CCL5 (10 ng/mL, R&D), or MVC (500 nmol/L, Selleckchem, S2003), followed by TMZ (500 μmol/L) treatment 4 h later. Apoptotic analysis was performed using Annexin-V Apoptosis Detection Kit (Invitrogen, 88-8007-74) according to the manufacturer’s instructions. Cell survival analysis was performed using CCK-8 (Dojindo, CK04) as previously described.^{48 (link)}

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Chemokine and Inhibitor Protocol

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Recombinant human CCL5 (cat# 278-RN), CXCL12 (cat# 350-NS) and pertussis toxin (PTX, cat# 3097) were purchased from R&D (Minneapolis, MN). Wortmannin (cat# W1628), 1-[6-[((17β)-3-Methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dione (U73122, cat# U6756) and retinoic acid (RA, cat# R2625) were from Sigma-Aldrich (St. Louis, MO).

Dedoni S., Campbell L.A., Harvey B.K., Avdoshina V, & Mocchetti I. (2018). The Orphan G Protein-coupled Receptor 75 Signaling is Activated by the Chemokine CCL5. Journal of neurochemistry, 146(5), 526-539.

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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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CoCl2 and SIN-1 Induced Cellular Responses

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CoCl₂ and SIN-1 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Recombinant human CCL5 was purchased from R&D Systems (Minneapolis, MN, USA).

Kim J.Y., Shin K.K., Lee A.L., Kim Y.S., Park H.J., Park Y.K., Bae Y.C, & Jung J.S. (2014). MicroRNA-302 induces proliferation and inhibits oxidant-induced cell death in human adipose tissue-derived mesenchymal stem cells. Cell Death & Disease, 5(8), e1385-.

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