Overview

  • Product name

  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    spleen 8 5.8%
    Inter-assay
    Sample n Mean SD CV%
    spleen 3 7.1%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    6.9 pg/ml
  • Range

    11.7 pg/ml - 750 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98 95% - 100%
    Serum 90 83% - 94%
    Hep Plasma 94 90% - 98%
    EDTA Plasma 96 88% - 101%
    Cit plasma 99 92% - 106%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Rat
  • Product overview

    CCL4 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of CCL4 protein in ratserum, plasma, and cell culture supernatants.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    CCL4 (Chemokine (C-C motif) ligand 4), also known as MIP-1 beta, is a protein which in rat is encoded by the MIP-1 beta gene. It is a CC chemokine with specificity for CCR5 receptors. It is a chemoattractant for natural killer cells, monocytes and a variety of other immune cells. MIP-1 beta is a major HIV-suppressive factor produced by CD8+ T cells. Performing-low memory CD8+ T cells that normally synthesize MIP-1-beta. By analysis of somatic cell hybrids and by in situ hybridization, the MIP-1 beta gene was assigned to 17q21-q23. Based on functional analysis of MIP-1β, one or more additional β chemokine receptors that transduce MIP-1β signals must also exist. MIP-1α and MIP-1β have been shown to lack affinity for the erythrocyte surface chemokine receptor/Duffy antigen that binds many α as well as β chemokines.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Rat CCL4 Capture Antibody 1 x 600µl
    10X Rat CCL4 Detector Antibody 2 vials
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BR 1 x 6ml
    Plate Seals 1 unit
    Rat CCL4 Lyophilized Recombinant Protein 1 x 600µl
    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

  • Function

    Monokine with inflammatory and chemokinetic properties. Binds to CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant MIP-1-beta induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV). The processed form MIP-1-beta(3-69) retains the abilities to induce down-modulation of surface expression of the chemokine receptor CCR5 and to inhibit the CCR5-mediated entry of HIV-1 in T-cells. MIP-1-beta(3-69) is also a ligand for CCR1 and CCR2 isoform B.
  • Sequence similarities

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

    N-terminal processed form MIP-1-beta(3-69) is produced by proteolytic cleavage after secretion from peripheral blood lymphocytes.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • MIP 1 beta
    • Secreted protein G 26
    • ACT 2
    • ACT-2
    • ACT2
    • AT744.1
    • AT744.2
    • C C motif chemokine 4
    • C C motif chemokine 4 like
    • C C motif chemokine ligand 4 like 1
    • C C motif chemokine ligand 4 like 2
    • CC chemokine ligand 4
    • CC chemokine ligand 4L1
    • CC chemokine ligand 4L1d2
    • CC chemokine ligand 4L2
    • CCL4
    • CCL4_HUMAN
    • CCL4L
    • ccl4l 1
    • CCL4L1
    • Chemokine (C C motif) ligand 4
    • Chemokine (C C motif) ligand 4 like 1
    • Chemokine (C C motif) ligand 4 like 1, telomeric
    • Chemokine (C C motif) ligand 4 like 2
    • Chemokine CC Motif Ligand 4
    • G 26
    • G 26 T lymphocyte secreted protein
    • G-26 T-lymphocyte-secreted protein
    • HC21
    • Immune activation 2
    • LAG 1
    • LAG-1
    • LAG1
    • Lymphocyte activation gene 1
    • Lymphocyte activation gene 1 protein
    • Macrophage inflammatory protein 1 beta
    • Macrophage inflammatory protein 1-beta
    • Macrophage inflammatory protein 1b2
    • MGC104418
    • MGC126025
    • MGC126026
    • MIP-1-beta
    • MIP-1-beta(1-69)
    • MIP-1-beta(3-69)
    • MIP1 beta
    • MIP1B
    • MIP1B1
    • Monocyte adherence induced protein 5 alpha
    • PAT 744
    • Protein H400
    • SCYA2
    • SCYA4
    • SCYA4L
    • SCYA4L1
    • SCYA4L2
    • SCYQ4L2
    • Secreted protein G 26
    • Secreted protein G26
    • SIS gamma
    • SIS-gamma
    • Small inducible cytokine A4
    • Small inducible cytokine A4 (homologous to mouse Mip 1b)
    • small inducible cytokine A4-like
    • Small-inducible cytokine A4
    • T cell activation protein 2
    • T-cell activation protein 2
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab240680 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • The CCL4 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.
  • The concentrations of CCL4 were measured in duplicates, interpolated from the CCL4 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (citrate) 50%, plasma (heparin) 50% and plasma (EDTA) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
  • Rat PBMCS were cultured in RPMI supplemented with 10% fetal calf serum. Cells were cultured for days in the presence or absence of PHA. Rat Spleen was cultured in the presence or absence of PHA and rat IL2 for 6 days. The concentrations of CCL4 were measured in duplicate and interpolated from the CCL4 standard curve and corrected for sample dilution. Undiluted samples are as follows: 3% PBMC culture supernatant and 25% spleen culture supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CCL4 concentration was determined to be 5,330 pg/mL in stimulated PBMC, 720 pg/mL in unstimulated PBMC, and 2,051 pg/mL in stimulated spleen.

Protocols

References

ab240680 has not yet been referenced specifically in any publications.

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