Overview

  • Product name

    Human CCL18 ELISA Kit
    See all CCL18 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 3%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 7.7%
  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.55 pg/ml
  • Range

    1.53 pg/ml - 100 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 92 87% - 95%
    Urine 90 86% - 94%
    Serum 90 85% - 93%
    Hep Plasma 99 92% - 102%
    EDTA Plasma 93 87% - 96%
    Cit plasma 103 97% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Cow
  • Product overview

    CCL18 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of human CCL18 protein in human serum, plasma, urine, synovial fluid and cell culture supernatant 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.

  • Notes

    Human CCL18 (macrophage inflammatory protein-4) belongs to the C-C motif containing chemokine family and contains a 20 amino acid (aa) signal peptide and a 69 aa residue mature protein. CCL18 was identified as a chemotactic factor for naïve T-cells, CD4+ and CD8+ T-cells, and nonactivated lymphocytes, suggesting that it plays an important role in immune responses. Additionally, MIP4 is mainly induced by Th2 type cytokines, such as IL-4 and IL-13, and inhibited by IFN-γ. High expression of MIP4 is seen in lung, lymph nodes, placenta, and macrophages derived from peripheral blood monocytes. Additionally, MIP4 is found in high levels in serum, plasma, and synovial fluid and in low levels in urine. Currently, a rodent homolog has not been identified.

  • 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 CCL18 Capture Antibody 1 x 600µl
    10X Human CCL18 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human CCL18 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

  • Function

    Chemotactic factor that attracts lymphocytes but not monocytes or granulocytes. May be involved in B-cell migration into B-cell follicles in lymph nodes. Attracts naive T-lymphocytes toward dendritic cells and activated macrophages in lymph nodes, has chemotactic activity for naive T-cells, CD4+ and CD8+ T-cells and thus may play a role in both humoral and cell-mediated immunity responses.
  • Tissue specificity

    Expressed at high levels in lung, lymph nodes, placenta, bone marrow, dendritic cells present in germinal centers and T-cell areas of secondary lymphoid organs and macrophages derived from peripheral blood monocytes. Not expressed by peripheral blood monocytes and a monocyte-to-macrophage differentiation is a prerequisite for expression. Expressed in synovial fluids from patients with rheumatoid and septic arthritis and in ovarian carcinoma ascitic fluid.
  • Sequence similarities

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

    Secreted.
  • Information by UniProt
  • Alternative names

    • Alternative macrophage activation associated CC chemokine 1
    • Alternative macrophage activation-associated CC chemokine 1
    • AMAC-1
    • AMAC1
    • CC chemokine PARC
    • CCL18
    • CCL18(4-69)
    • CCL18_HUMAN
    • CKb7
    • DC-CK1
    • DCCK1
    • Dendritic cell chemokine 1
    • Macrophage inflammatory protein 4
    • MIP-4
    • MIP4
    • PARC
    • Pulmonary and activation regulated chemokine
    • Pulmonary and activation-regulated chemokine
    • SCYA18
    • Small inducible cytokine A18
    • Small-inducible cytokine A18
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab211649 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

  • Standard curve comparison between human MIP4 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 6-fold increase in sensitivity.

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

  • 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 concentrations of MIP4 were measured in duplicates, interpolated from the MIP4 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:1,200, plasma (citrate) 1:600, plasma (EDTA) 1:600, and plasma (heparin) 1:1,200. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP4 concentration was determined to be 79,615 pg/mL in serum, 57,039 pg/mL in plasma (citrate), 59,254 pg/mL in plasma (EDTA) and 71,426 pg/mL in plasma (heparin).

  • The concentrations of MIP4 were measured in duplicates, interpolated from the MIP4 standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 50%, synovial fluid 1:600, PBMC stimulated media (Donor 1) 6.25% and PBMC stimulated media (Donor 2) 3.13%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP4 concentration was determined to be 34.5 pg/mL in urine, 57,576 pg/mL in synovial fluid, 1,251 pg/mL in PBMC stimulated media (Donor 1), and 2,448 pg/mL in PBMC stimulated media (Donor 2). PBMC samples were cultured in RPMI media with 10% fetal bovine serum and 1% PenStrep and stimulated with 1.5% PHA-M for 48 hours.

  • All PBMC media samples were diluted to 3.13% and analyzed. The concentrations of MIP4 were measured in duplicates and interpolated from the MIP4 standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP4 concentration was determined to be 46 pg/mL in unstimulated PBMC Media (Donor 1), 1,292 pg/mL in stimulated PBMC Media (Donor 1), 87 pg/mL in PBMC unstimulated media (Donor 2), and 2,521 pg/mL in PBMC stimulated media (Donor 2). PBMC samples were cultured in RPMI media with 10% fetal bovine serum and 1% PenStrep (unstimulated samples). Then PBMC samples were stimulated with 1.5% PHA-M for 48 hours.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP4 concentration was determined to be 47,858 pg/mL with a range of 29,081 – 66,007 pg/mL.

Protocols

References

ab211649 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab211649.
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