Overview

  • Product name

  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human serum 3 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 5 7%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Other biological fluids, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    27.1 pg/ml
  • Range

    89.4 pg/ml - 1500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 104 97% - 110%
    Serum 92 90% - 95%
    Cell culture extracts 106 100% - 115%
    Other biological fluids 117 116% - 117%
    Tissue Extracts 108 91% - 121%
    Heparin Plasma 108 102% - 112%
    EDTA Plasma 107 102% - 115%
    Citrate Plasma 103 91% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    MIP5 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MIP5 protein in human serum, plasma, cell culture supernatant, synovial fluid, and cell and tissue extract samples.


    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.


    Sensitivity:


    Samples in 1X Cell Extraction Buffer PTR - 30.6 pg/mL


    Samples in Sample Diluent NS - 27.1 pg/mL

  • Notes

    MIP5 is a secreted monomeric chemotactic factor that attracts T-cells and monocytes, but not neutrophils, eosinophils, or B-cells. MIP5 acts mainly via CC chemokine receptor CCR1. It also binds to CCR3. MIP5 can be cleaved into 3 chains, MIP5(22-92), MIP5(25-92) and MIP5(29-92). These cleavage products are more potent chemo-attractants than MIP5. They are also found in high levels in synovial fluids from rheumatoid patients.

  • 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 Human MIP5 Capture Antibody 1 x 600µl
    10X Human MIP5 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BI 1 x 6ml
    Human MIP5 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 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

    Chemotactic factor that attracts T-cells and monocytes, but not neutrophils, eosinophils, or B-cells. Acts mainly via CC chemokine receptor CCR1. Also binds to CCR3. CCL15(22-92), CCL15(25-92) and CCL15(29-92) are more potent chemoattractants than the small-inducible cytokine A15.
  • Tissue specificity

    Most abundant in heart, skeletal muscle and adrenal gland. Lower levels in placenta, liver, pancreas and bone marrow. CCL15(22-92), CCL15(25-92) and CCL15(29-92) are found in high levels in synovial fluids from rheumatoid patients.
  • Sequence similarities

    Belongs to the intercrine beta (chemokine CC) family.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • C-C motif chemokine 15
    • C-C motif chemokine ligand 15
    • CC chemokine 3
    • CCL15
    • CCL15(29-92)
    • CCL15_HUMAN
    • Chemokine (C C motif) ligand 15
    • Chemokine CC-2
    • Chemokine CC2
    • HCC-2
    • HCC2
    • HMRP 2B
    • HMRP2B
    • Leukotactin 1
    • Leukotactin-1
    • LKN-1
    • LKN1
    • Macrophage inflammatory protein 5
    • MIP 1D
    • MIP-1 delta
    • MIP-1D
    • MIP-5
    • MIP1 delta
    • MIP1d
    • MIP5
    • MRP 2B
    • Mrp-2b
    • Mrp2b
    • NCC 3
    • NCC-3
    • NCC3
    • New CC chemokine 3
    • OTTHUMP00000163955
    • SCYA15
    • SCYL3
    • Small inducible cytokine A15
    • Small inducible cytokine subfamily A (Cys Cys), member 15
    • Small-inducible cytokine A15
    • SY15
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab217774 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.

     

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

  • The MIP5 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 MIP5 were measured in duplicates, interpolated from the MIP5 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 20%, plasma (citrate) 20%, plasma (heparin) 20% and plasma (EDTA) 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP5 concentration was determined to be 8705 pg/mL in neat serum, 7196 pg/mL in neat plasma (citrate), 6848 pg/mL in neat plasma (heparin), and 7231 pg/mL in neat plasma (EDTA).

  • The concentrations of MIP5 were measured in duplicates, interpolated from the MIP5 standard curves and corrected for sample dilution. Undiluted samples are as follows: HepG2 cell culture supernatant 100% (neat), and synovial fluid 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP5 concentration was determined to be 706.2 pg/mL in neat HepG2 cell culture supernatant and 5,385 pg/mL in neat synovial fluid.

  • The concentrations of MIP5 were measured in duplicate and interpolated from the MIP5 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP5 concentration was determined to be 1293 pg/mL in HT-29 cell extract and 685.5 pg/mL in colon tissue extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP5 concentration was determined to be 7978 pg/mL with a range of 3361 – 13894 pg/mL.

  • Serum was pooled from fifty apparently healthy donors and compared to serum from a patient who tested positive for rheumatoid factor (RF value 406.2 IU/mL). The concentrations of MIP5 were measured from serum samples in 3 different dilutions in duplicates and interpolated from the MIP5 standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean MIP5 concentration was determined to be 8753 pg/mL in healthy human serum and 26193 pg/mL in rheumatoid arthritis patient serum.

Protocols

References

ab217774 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab217774.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up