Overview

  • Product name

    Human MIP1b ELISA Kit
    See all CCL4/MIP-1 beta kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 3.9%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 2.7%
  • Sample type

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.96 pg/ml
  • Range

    4.69 pg/ml - 300 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 100 98% - 102%
    Serum 98 96% - 99%
    Heparin Plasma 100 98% - 101%
    EDTA Plasma 98 96% - 101%
    Citrate Plasma 101 97% - 104%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    Human MIP1b SimpleStep ELISA kit (ab178010) has been re-developed with new capture and detector antibodies. This new kit has the same name but a different product number (ab216173). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying IL-17A in human serum, plasma and cell culture supernatants.


     


    MIP1b in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MIP1b protein in human serum, plasmas, and cell culture supernatant. 


    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 substrate 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.

  • Notes

    MIP-1-beta (CCL4) is a monokine with inflammatory and chemokinetic properties. MIP-1-beta binds to CCR5 and is 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. Structurally, MIP-1-beta forms a heterodimer with MIP-1-alpha (4-69) and is secreted from peripheral blood lymphocytes.

  • 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 Wash Buffer PT (ab206977) 1 x 20ml
    20X Human MIP1b Capture Antibody 1 x 300µl
    20X Human MIP1b Detector Antibody 1 x 300µl
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Human MIP1b Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    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

Applications

Our Abpromise guarantee covers the use of ab216173 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.

     

  • Standard curve comparison between human MIP1b SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 3-fold increase in sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of MIP1b were measured in duplicate, interpolated from the MIP1b standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 100%, plasma (heparin) 50% and stimulated PBMC supernatant 0.05%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP1b concentration was determined to be 159 pg/mL in serum, 147 pg/mL in plasma (citrate), 122 pg/mL in plasma (EDTA), 147 pg/mL in plasma (heparin), and 168 pg/mL in stimulated PBMC supernatant.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP1b concentration was determined to be 66 pg/mL with a range of 26 – 122 pg/mL.

  • PBMCs were treated with 1% PHA-M for 4 days.

Protocols

References

ab216173 has not yet been referenced specifically in any publications.

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