Human CCL5/RANTES Antibody

Catalog # Availability Size / Price Qty
AF-278-NA
AF-278-SP
Detection of Recombinant Human and Mouse CCL5/RANTES by Western Blot.
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Product Details
Citations (30)
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Human CCL5/RANTES Antibody Summary

Species Reactivity
Human
Specificity
Detects human CCL5/RANTES in direct ELISAs. Detects human CCL5/RANTES and mouse CCL5/RANTES Western blots. In direct ELISAs, approximately 40% cross-reactivity with recombinant canine CCL5/RANTES is observed and approximately 20% cross-reactivity with recombinant cotton rat CCL5/RANTES is observed. In Western blots, no cross-reactivity with recombinant human (rh) CCL3 and rhCCL4 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human CCL5/RANTES
Ser24-Ser91
Accession # P13501
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
Immersion fixed paraffin-embedded sections of human lymphoma and human tonsil
Immunocytochemistry
5-15 µg/mL
See below
Neutralization
Measured by its ability to neutralize CCL5/RANTES-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 0.1-0.4 µg/mL in the presence of 0.01 µg/mL Recombinant Human CCL5/RANTES.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Recombinant Human and Mouse CCL5/RANTES antibody by Western Blot. View Larger

Detection of Recombinant Human and Mouse CCL5/RANTES by Western Blot. Western blot shows 25 ng of Recombinant Human CCL5/RANTES (278-RN), Recombinant Mouse CCL5/RANTES (478-MR), Recombinant Human CCL3/MIP-1a Isoform LD78a (270-LD), and Recombinant Human CCL4/MIP-1 beta (271-BME). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-278-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for CCL5/RANTES at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Immunocytochemistry CCL5/RANTES antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

CCL5/RANTES in Human PBMCs. CCL5/RANTES was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-278-NA) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (orange; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Immunohistochemistry CCL5/RANTES antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

CCL5/RANTES in Human Tonsil. CCL5/RANTES was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-278-NA) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western Detection of Human CCL5/RANTES antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human CCL5/RANTES by Simple WesternTM. Simple Western lane view shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200nM PMA for 24 hrs and 10ug/mL LPS for 3 hrs, loaded at 0.2 mg/mL. A specific band was detected for CCL5/RANTES at approximately 7 kDa (as indicated) using 10 µg/mL of Goat Anti-Human CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-278-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Neutralization Chemotaxis Induced by CCL5/RANTES and Neutralization by Human CCL5/RANTES Antibody. View Larger

Chemotaxis Induced by CCL5/RANTES and Neutralization by Human CCL5/RANTES Antibody. Recombinant Human CCL5/RANTES (Catalog # 278-RN) chemoattracts the BaF3 mouse pro-B cell line transfected with human CCR5 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (AR002). Chemotaxis elicited by Recombinant Human CCL5/RANTES (0.01 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-278-NA). The ND50 is typically 0.1‑0.4 µg/mL.

Immunohistochemistry View Larger

Detection of CCL5/RANTES in Human Tonsil. CCL5/RANTES was detected in immersion fixed paraffin-embedded sections of Human Tonsil using Goat Anti-Human CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-278-NA) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Immunohistochemistry Detection of Human CCL5/RANTES by Immunohistochemistry View Larger

Detection of Human CCL5/RANTES by Immunohistochemistry CCL5-CCR5 axis induced aerobic glycolysis by regulation of AMPK signaling(A) Western blot for AMPK, c-Myc, HIF-1 alpha and Akt in breast cancer cells co-cultured with 15mM lactic acid-activated THP-1 macrophages (ratio 1:1) for 72 h. Results presented were representatives of at least three independent experiments. (B) The expression of AMPK downstream signaling target ACC in breast cancer cells co-cultured as in (A). (C) MDA-MB-231 and MCF-7 cells were transfected with 50 nM AMPK alpha 1 siRNA, or pretreated with 10μM compound C for 4 h, and then incubated with 15mM lactic acid-activated THP-1 macrophages (ratio 1:1) for 48 h. The glucose uptake, lactic acid production and ATP levels were detected. (D) The inhibition of AMPK abrogated macrophage-induced EMT in MCF-7 cells. Cells were treated as described in (C). After co-culture, the expression of EMT markers, E-cadherin and vimentin, was measured by western blot. (E) Recombinant human CCL5 induced the phosphorylation of AMPK in MDA-MB-231 and MCF-7/CCR5 cells. 106 cells were treated with 50ng/ml CCL5 for defferent time points as indicated, and phosphorylated AMPK and total AMPK were investigated by western blot. (F) Inhibition of CCR5 in MDA-MB-231 cells significantly attenuated macrophage-induced AMPK phosphorylation. MDA-MB-231 cells were transfected with shRNAs designed against CCR5, or pre-treated with 5μM Maraviroc for 2 h, then co-cultured with 15 mM lactate-activated macrophages as described in (A). After co-culture, the phosphorylation of AMPK was detected by western blot. (G) Expressions of CCL5, CCR5 and p-AMPK in samples obtained from breast cancer patients (n =28). Scale bars represent 50 μm. *, P<0.05; **, P<0.01. Image collected and cropped by CiteAb from the following open publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.22786), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human CCL5/RANTES by Western Blot View Larger

Detection of Human CCL5/RANTES by Western Blot Lactate-activated macrophages induced glycolysis through CCL5-CCR5 axis(A) Glucose uptake, lactic acid production and ATP levels in breast cancer cells co-cultured with lactate-activated THP-1 macrophages, with or without 5μg/ml anti-CCL5 neutralizing antibody. The co-culture system was described in Figure 5C. (B) Western blots for glycolytic enzymes in breast cancer cells treated as in (A). (C) MDA-MB-231 cells were transfected with shRNAs designed against CCR5, or pre-treated with 5μM Maraviroc for 2 h, and then subjected to cell co-culture. Glucose uptake, lactic acid production and ATP levels were measured after co-culture. The co-culture system was described in Figure 5C. (D) The protein levels of HK2, PKM2 and LDHA in MDA-MB-231 cells cultured as in (C). (E) Recombinant human CCL5 induced aerobic glycolysis in breast cancer cells. MDA-MB-231 and MCF-7/CCR5 cells were treated with increasing concentrations of CCL5 for 12 h, and glucose uptake, lactic acid production and ATP levels were detected. (F) Western blots for glycolytic enzymes in MDA-MB-231 and MCF-7/CCR5 cells after stimulation with CCL5. *, P<0.05; **, P<0.01. Image collected and cropped by CiteAb from the following open publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.22786), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CCL5/RANTES

RANTES (Regulated upon Activation, Normal T cell Expressed and presumably Secreted) is a member of the beta (C-C) chemokine subfamily and is now designated CCL5. It binds and activates the chemokine receptors CCR1, 3 and 5.

Entrez Gene IDs
6352 (Human); 20304 (Mouse); 403522 (Canine); 493689 (Feline)
Alternate Names
CCL5; chemokine (C-C motif) ligand 5; D17S136Enormally T-expressed, and presumably secreted; EoCP; Eosinophil chemotactic cytokine; RANTES; SISd; SIS-delta; small inducible cytokine A5 (RANTES); small inducible cytokine subfamily A (Cys-Cys), member 5; Small-inducible cytokine A5; T cell-specific protein P228; T-cell specific protein p288; TCP228T-cell-specific protein RANTES

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Citations for Human CCL5/RANTES Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

30 Citations: Showing 1 - 10
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  1. Controlled release of biological factors for endogenous progenitor cell migration and intervertebral disc extracellular matrix remodelling
    Authors: Leslie Frapin, Johann Clouet, Claire Chédeville, Constantin Moraru, Edouard Samarut, Nina Henry et al.
    Biomaterials
  2. Preterm Birth Is Associated With Immune Dysregulation Which Persists in Infants Exposed to Histologic Chorioamnionitis
    Authors: Gemma Sullivan, Paola Galdi, Nis Borbye-Lorenzen, David Q. Stoye, Gillian J. Lamb, Margaret J. Evans et al.
    Frontiers in Immunology
  3. PARP inhibitors promote stromal fibroblast activation by enhancing CCL5 autocrine signaling in ovarian cancer
    Authors: Xiaoting Li, Tian Fang, Sen Xu, Ping Jin, Dongchen Zhou, Zhengzheng Wang et al.
    npj Precision Oncology
  4. Blockade of Autocrine CCL5 Responses Inhibits Zika Virus Persistence and Spread in Human Brain Microvascular Endothelial Cells
    Authors: MC Mladinich, JN Conde, WR Schutt, SY Sohn, ER Mackow
    MBio, 2021-08-17;12(4):e0196221.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. The Protein Kinase Receptor Modulates the Innate Immune Response against Tacaribe Virus
    Authors: H Moreno, S Kunz
    Viruses, 2021-07-07;13(7):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. Dissecting spatial heterogeneity and the immune-evasion mechanism of CTCs by single-cell RNA-seq in hepatocellular carcinoma
    Authors: YF Sun, L Wu, SP Liu, MM Jiang, B Hu, KQ Zhou, W Guo, Y Xu, Y Zhong, XR Zhou, ZF Zhang, G Liu, S Liu, YH Shi, Y Ji, M Du, NN Li, GB Li, ZK Zhao, XY Huang, LQ Xu, QC Yu, DH Peng, SJ Qiu, HC Sun, M Dean, XD Wang, WY Chung, AR Dennison, J Zhou, Y Hou, J Fan, XR Yang
    Nature Communications, 2021-07-02;12(1):4091.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  7. CCR5/CCL5 axis interaction promotes migratory and invasiveness of pancreatic cancer cells
    Authors: SK Singh, MK Mishra, IA Eltoum, S Bae, JW Lillard, R Singh
    Sci Rep, 2018-01-22;8(1):1323.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  8. An initial investigation into endothelial CC chemokine expression in the human rheumatoid synovium.
    Authors: Rump L, Mattey D, Kehoe O, Middleton J
    Cytokine, 2017-09-01;97(0):133-140.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  9. The Ovarian Cancer Chemokine Landscape Is Conducive to Homing of Vaccine-Primed and CD3/CD28-Costimulated T Cells Prepared for Adoptive Therapy.
    Authors: Zsiros E, Duttagupta P, Dangaj D, Li H, Frank R, Garrabrant T, Hagemann I, Levine B, June C, Zhang L, Wang E, Marincola F, Bedognetti D, Powell D, Tanyi J, Feldman M, Kandalaft L, Coukos G
    Clin Cancer Res, 2015-02-23;21(12):2840-50.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  10. NK cell responses to simian immunodeficiency virus vaginal exposure in naive and vaccinated rhesus macaques.
    Authors: Shang L, Smith A, Duan L, Perkey K, Qu L, Wietgrefe S, Zupancic M, Southern P, Masek-Hammerman K, Reeves R, Johnson R, Haase A
    J Immunol, 2014-06-04;193(1):277-84.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Types: Whole Tissue
    Applications: IHC
  11. Lenalidomide inhibits lymphangiogenesis in preclinical models of mantle cell lymphoma.
    Authors: Song K, Herzog B, Sheng M, Fu J, McDaniel J, Chen H, Ruan J, Xia L
    Cancer Res, 2013-10-24;73(24):7254-64.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  12. Expression and regulation of chemokines in murine and human type 1 diabetes.
    Authors: Sarkar SA, Lee CE, Victorino F, Nguyen TT, Walters JA, Burrack A, Eberlein J, Hildemann SK, Homann D
    Diabetes, 2011-12-30;61(2):436-46.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  13. Irritant-induced migration of Langerhans cells coincides with an IL-10-dependent switch to a macrophage-like phenotype.
    Authors: Ouwehand K, Oosterhoff D, Breetveld M
    J. Invest. Dermatol., 2010-11-11;131(2):418-25.
    Species: Human
    Sample Types: Whole Tissue
    Applications: Neutralization
  14. Comprehensive assessment of chemokine expression profiles by flow cytometry.
    Authors: Eberlein J, Nguyen TT, Victorino F, Golden-Mason L, Rosen HR, Homann D
    J. Clin. Invest., 2010-02-08;120(3):907-23.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  15. Villitis of unknown etiology is associated with a distinct pattern of chemokine up-regulation in the feto-maternal and placental compartments: implications for conjoint maternal allograft rejection and maternal anti-fetal graft-versus-host disease.
    Authors: Kim MJ, Romero R, Kim CJ, Tarca AL, Chhauy S, LaJeunesse C, Lee DC, Draghici S, Gotsch F, Kusanovic JP, Hassan SS, Kim JS
    J. Immunol., 2009-03-15;182(6):3919-27.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  16. Tumour necrosis factor alpha stimulates the production of monocyte chemoattractants by extravillous trophoblast cells via differential activation of MAPK pathways.
    Authors: Renaud SJ, Sullivan R, Graham CH
    Placenta, 2009-02-08;30(4):313-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  17. Disrupting functional interactions between platelet chemokines inhibits atherosclerosis in hyperlipidemic mice.
    Authors: Koenen RR, von Hundelshausen P, Nesmelova IV, Zernecke A, Liehn EA, Sarabi A, Kramp BK, Piccinini AM, Paludan SR, Kowalska MA, Kungl AJ, Hackeng TM, Mayo KH, Weber C
    Nat. Med., 2009-01-04;15(1):97-103.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  18. Altered levels of CC chemokines during pulmonary CMV predict BOS and mortality post-lung transplantation.
    Authors: Weigt SS, Elashoff RM, Keane MP, Strieter RM, Gomperts BN, Xue YY, Ardehali A, Gregson AL, Kubak B, Fishbein MC, Saggar R, Ross DJ, Lynch JP, Zisman DA, Belperio JA
    Am. J. Transplant., 2008-07-01;8(7):1512-22.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  19. Expanded-polyglutamine huntingtin protein suppresses the secretion and production of a chemokine (CCL5/RANTES) by astrocytes.
    Authors: Chou SY, Weng JY, Lai HL, Liao F, Sun SH, Tu PH, Dickson DW, Chern Y
    J. Neurosci., 2008-03-26;28(13):3277-90.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  20. Mesenchymal stem cells within tumour stroma promote breast cancer metastasis.
    Authors: Karnoub AE, Dash AB, Vo AP, Sullivan A, Brooks MW, Bell GW, Richardson AL, Polyak K, Tubo R, Weinberg RA
    Nature, 2007-10-04;449(7162):557-63.
    Species: Human
    Sample Types: In Vivo
    Applications: Neutralization
  21. Renal transplantation: examination of the regulation of chemokine binding during acute rejection.
    Authors: Ali S, Malik G, Burns A, Robertson H, Kirby JA
    Transplantation, 2005-03-27;79(6):672-9.
    Species: Human
    Sample Types: Whole Cells, Whole Tissue
    Applications: ICC, IHC-P
  22. The beta chemokines CCL4 and CCL5 enhance adhesion of specific CD4+ T cell subsets to human brain endothelial cells.
    Authors: Quandt J, Dorovini-Zis K
    J. Neuropathol. Exp. Neurol., 2004-04-01;63(4):350-62.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  23. Phenotypic and functional analysis of T cells homing into the CSF of subjects with inflammatory diseases of the CNS.
    Authors: Giunti D, Borsellino G, Benelli R, Marchese M, Capello E, Valle MT, Pedemonte E, Noonan D, Albini A, Bernardi G, Mancardi GL, Battistini L, Uccelli A
    J. Leukoc. Biol., 2003-05-01;73(5):584-90.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  24. Simian immunodeficiency viruses with defective nef genes show increased susceptibility to the noncytotoxic antiviral activity of CD8+ lymphocytes.
    Authors: Binninger-Schinzel D, 2019, Norley S, Adler HS, Oberg HH, Kurth R
    2002-03-01;294(1):209-21.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  25. Bleomycin stimulates lung fibroblast and epithelial cell lines to release eosinophil chemotactic activity.
    Authors: Sato E, Koyama S
    Eur. Respir. J., 2000-11-01;16(5):951-8.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  26. Lactate-activated macrophages induced aerobic glycolysis and epithelial-mesenchymal transition in breast cancer by regulation of CCL5-CCR5 axis: a positive metabolic feedback loop
    Authors: Sensen Lin, Li Sun, Xiaodan Lyu, Xiongfei Ai, Danyu Du, Nan Su et al.
    Oncotarget
  27. Titanium salts tested in reconstructed human skin with integrated MUTZ ‐3‐derived Langerhans cells show an irritant rather than a sensitizing potential
    Authors: Charlotte T. Rodrigues Neves, Sander W. Spiekstra, Niels P. J. de Graaf, Thomas Rustemeyer, Albert J. Feilzer, Cees J. Kleverlaan et al.
    Contact Dermatitis
  28. Human Bone Marrow Mesenchymal Stem Cells Induce Collagen Production and Tongue Cancer Invasion
    Authors: Sirpa Salo, Carolina Bitu, Kalle Merkku, Pia Nyberg, Ibrahim O Bello, Jussi Vuoristo et al.
    PLoS ONE
  29. Toll-Like Receptor 3 as a Recurrence Risk Factor and a Potential Molecular Therapeutic Target in Colorectal Cancer
    Authors: Yoshida T, Miura T, Matsumiya T et al.
    Clin Exp Gastroenterol
  30. Low CCR5 expression protects HIV-specific CD4+ T cells of elite controllers from viral entry
    Authors: Mathieu Claireaux, Rémy Robinot, Jérôme Kervevan, Mandar Patgaonkar, Isabelle Staropoli, Anne Brelot et al.
    Nature Communications

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