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    products/elisa/human-rantes-elisa-kit-ccl5-ab174446.pdf

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Immunology Secreted Molecules CCN Family
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RecombinantSimpleStep

Human RANTES ELISA Kit (CCL5) (ab174446)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2) Submit a question References (7)

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Other - Human RANTES ELISA Kit (CCL5) (ab174446)
  • Human RANTES standard curve comparison.
  • Example of human RANTES standard curve in Sample Diluent NS.
  • Interpolated concentrations of native RANTES in human serum, and plasma samples.
  • Interpolated concentrations of native RANTES in human PBMC cell culture supernatant stimulated with 1.5% PHA-M for 36 hours.
  • Serum from ten individual healthy human male donors was measured in duplicate.
  • Comparison of PHA-M stimulated versus unstimulated PBMC supernatant.
  • Sandwich ELISA - Human RANTES ELISA Kit (CCL5) (ab174446)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 0.091 pg/ml
  • Range: 0.94 pg/ml - 60 pg/ml
  • Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human RANTES ELISA Kit (CCL5)
    See all RANTES kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 2.8%
  • Sample type

    Cell culture supernatant, Serum, Plasma, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.091 pg/ml
  • Range

    0.94 pg/ml - 60 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 104 95% - 114%
    Serum 107 77% - 132%
    Hep Plasma 91 79% - 100%
    EDTA Plasma 105 81% - 119%
    Cit plasma 105 82% - 124%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human RANTES ELISA kit (CCL5) (ab174446) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of RANTES protein in human plasma, serum and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate human RANTES with 0.091 pg/ml sensitivity.

    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

            -Single-wash protocol reduces assay time to 90 minutes or less
            -High sensitivity, specificity and reproducibility from superior antibodies
            -Fully validated in biological samples
            -96-wells plate breakable into 12 x 8 wells strips

    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.

  • Notes

    RANTES (Regulated on Activation, Normal T cell Expressed and Secreted) is a chemotactic cytokine for T cells, eosinophils, and basophils. RANTES also acts as a recruitment signal for leukocytes into inflammatory sites with the help of cyotokines (IL-2 and IFN-γ) released by CD8+ T cells. RANTES also activates and induces proliferation of natural-killer cells to form CHAK (CC-Chemokine-activated killer) cells. RANTES has also been identified to inhibit infection of HIV.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human RANTES Capture Antibody 1 x 600µl
    10X Human RANTES Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPR 1 x 6ml
    Human RANTES Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Immunology
    • Secreted Molecules
    • CCN Family
    • Immunology
    • Innate Immunity
    • Chemokines
    • Beta Chemokines (CC)
    • Immunology
    • Secreted Molecules
    • Other secreted molecules
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Chemokines
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
    • Immunology
    • Immune System Diseases
    • Antiviral Signaling
    • HIV-related
    • Neuroscience
    • Processes
  • Function

    Chemoattractant for blood monocytes, memory T-helper cells and eosinophils. Causes the release of histamine from basophils and activates eosinophils. Binds to CCR1, CCR3, CCR4 and CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant RANTES protein induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV). The processed form RANTES(3-68) acts as a natural chemotaxis inhibitor and is a more potent inhibitor of HIV-1-infection. The second processed form RANTES(4-68) exhibits reduced chemotactic and HIV-suppressive activity compared with RANTES(1-68) and RANTES(3-68) and is generated by an unidentified enzyme associated with monocytes and neutrophils.
  • Tissue specificity

    T-cell and macrophage specific.
  • Sequence similarities

    Belongs to the intercrine beta (chemokine CC) family.
  • Post-translational
    modifications

    N-terminal processed form RANTES(3-68) is produced by proteolytic cleavage, probably by DPP4, after secretion from peripheral blood leukocytes and cultured sarcoma cells.
    The identity of the O-linked saccharides at Ser-27 and Ser-28 are not reported in PubMed:1380064. They are assigned by similarity.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P13501 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Beta chemokine RANTES
    • Beta chemokine RANTES precursor
    • C C motif chemokine 5
    • CCL 5
    • CCL5
    • CCL5_HUMAN
    • Chemokine (C C motif) ligand 5
    • Chemokine CC Motif Ligand 5
    • D17S136E
    • EoCP
    • Eosinophil chemotactic cytokine
    • MGC17164
    • RANTES(4-68)
    • Regulated upon activation normally T expressed and presumably secreted
    • SCYA 5
    • SCYA5
    • SIS delta
    • SIS-delta
    • SISd
    • Small inducible cytokine A5
    • Small inducible cytokine A5 (RANTES)
    • Small inducible cytokine subfamily A (Cys Cys) member 5
    • Small-inducible cytokine A5
    • T cell specific protein p288
    • T cell specific protein RANTES
    • T cell specific RANTES protein
    • T cell-specific protein P228
    • T-cell-specific protein RANTES
    • TCP 228
    • TCP228
    see all
  • Database links

    • Entrez Gene: 6352 Human
    • Omim: 187011 Human
    • SwissProt: P13501 Human
    • Unigene: 514821 Human

    Associated products

    • Alternative Versions

      • Human RANTES Antibody Pair - BSA and Azide free (CCL5) (ab243971)

    Images

    • Other - Human RANTES ELISA Kit (CCL5) (ab174446)
      Other - Human RANTES ELISA Kit (CCL5) (ab174446)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Human RANTES standard curve comparison.
      Human RANTES standard curve comparison.

      Standard Curve comparison between human RANTES SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

    • Example of human RANTES standard curve in Sample Diluent NS.
      Example of human RANTES standard curve in Sample Diluent NS.

      The RANTES standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native RANTES in human serum, and plasma samples.
      Interpolated concentrations of native RANTES in human serum, and plasma samples.

      The concentrations of RANTES were measured in duplicates, interpolated from the RANTES standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 2.5%, plasma (citrate) 0.63%, plasma (heparin) 2.5% and plasma (EDTA) 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RANTES concentration was determined to be 2,141 pg/mL in neat serum, 13,050 pg/mL in plasma (citrate) and 3,157 pg/mL in plasma (heparin) and 4,457 pg/mL plasma (EDTA).

    • Interpolated concentrations of native RANTES in human PBMC cell culture supernatant stimulated with 1.5% PHA-M for 36 hours.
      Interpolated concentrations of native RANTES in human PBMC cell culture supernatant stimulated with 1.5% PHA-M for 36 hours.

      The supernatant was decanted and assayed. The concentrations of RANTES were measured in duplicates, interpolated from the RANTES standard curves and corrected for sample dilution. Undiluted samples are as follows: PBMC supernatant 1.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RANTES concentration was determined to be 2,156 pg/mL in neat PBMC supernatant.

    • Serum from ten individual healthy human male donors was measured in duplicate.
      Serum from ten individual healthy human male donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RANTES concentration was determined to be 977 pg/mL with a range of 135–2,378 pg/mL.

    • Comparison of PHA-M stimulated versus unstimulated PBMC supernatant.
      Comparison of PHA-M stimulated versus unstimulated PBMC supernatant.

      PBMCs were stimulated with 1.5% PHA-M for 36 hours. The concentrations of RANTES were measured in duplicates, interpolated from the RANTES standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RANTES concentration was determined to be 2,156 pg/mL in neat PBMC stimulated supernatant and non-detectable in unstimulated supernatant.

    • Sandwich ELISA - Human RANTES ELISA Kit (CCL5) (ab174446)
      Sandwich ELISA - Human RANTES ELISA Kit (CCL5) (ab174446)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab174446? Please let us know so that we can cite the reference in this datasheet.

    ab174446 has been referenced in 7 publications.

    • Jia J  et al. Cutaneous neurofibroma cells with active YAP promotes proliferation of macrophages resulting in increased accumulation of macrophages by modulating CCL5 and TGF-ß1. Oncol Rep 43:1319-1330 (2020). PubMed: 32323813
    • Li S  et al. Clinical and pathological investigation of patients with severe COVID-19. JCI Insight 5:N/A (2020). PubMed: 32427582
    • Hosseinkhani B  et al. (Sub)populations of extracellular vesicles released by TNF-a -triggered human endothelial cells promote vascular inflammation and monocyte migration. J Extracell Vesicles 9:1801153 (2020). PubMed: 32944190
    • Hosseinkhani B  et al. Extracellular Vesicles Work as a Functional Inflammatory Mediator Between Vascular Endothelial Cells and Immune Cells. Front Immunol 9:1789 (2018). PubMed: 30131806
    • Aznar MA  et al. CD137 (4-1BB) Costimulation Modifies DNA Methylation in CD8+ T Cell-Relevant Genes. Cancer Immunol Res 6:69-78 (2018). PubMed: 29133290
    • Li BH  et al. Steatosis induced CCL5 contributes to early-stage liver fibrosis in nonalcoholic fatty liver disease progress. Transl Res 180:103-117.e4 (2017). Sandwich ELISA ; Human . PubMed: 27639593
    • Lai XM  et al. HAF mediates the evasive resistance of anti-angiogenesis TKI through disrupting HIF-1a and HIF-2a balance in renal cell carcinoma. Oncotarget 8:49713-49724 (2017). Sandwich ELISA ; Human . PubMed: 28572533

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Abreview for ab174446 - Human RANTES ELISA Kit (CCL5)

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We measured the concentration of Human RANTES by using this Human RANTES SimpleStep ELISA® Kit. Human plasma were further diluted at 1:40, however the final optical density from some samples were still much more than the plate reader can be recorded. As far as the Abcam team was informed about that results, they immediately sent us another kit free of charge to repeat experiments. Hence, we further diluted human plasma at 1:200, the final optical density from all of samples were between the lowest and the highest standard. Quite perfect kit, and quite perfect service!
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 10 2022

    Abreview for Human RANTES SimpleStep ELISA® Kit

    Average Average 3/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We measured the concentration of Human RANTES by using this Human RANTES SimpleStep ELISA® Kit. Blood were collected in the vacutainer EDTA tubes, the upper layer of plasma was collected after centrifugation and frozen until determination. Samples were further diluted at 1:40 into Sample Diluent NS and assay according to the protocol, however the final optical density from some samples were still much more than the plate reader can be recorded.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 15 2021

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