Overview

  • Product name

    Human Lipocalin-2 ELISA Kit
    See all Lipocalin-2 / NGAL kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Plasma-EDTA 5 3.8%
    Inter-assay
    Sample n Mean SD CV%
    Plasma-EDTA 3 2.7%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    14.8 pg/ml
  • Range

    46.9 pg/ml - 3000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 104 101% - 106%
    Saliva 109 107% - 111%
    Urine 110 102% - 118%
    Serum 105 104% - 108%
    Heparin Plasma 103 99% - 107%
    EDTA Plasma 109 107% - 111%
    Citrate Plasma 106 104% - 109%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    Lipocalcin-2 in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of lipocalin-2 protein in human serum, plasma, cell culture supernatant, urine and saliva.


    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

    Lipocalin-2 (also known as Neutrophil gelatinase-associated lipocalin or NGAL) is an iron binding and iron trafficking protein. Lipocalin-2 is involved in multiple cellular processes including apoptosis, innate immunity and renal development. Mice deficient in Lipocalin-2 appear normal but have increased susceptibility to bacterial infection. The bacteriostatic function may be related to Lipocalin-2 limiting bacterial iron supply. Human Lipocalin-2 has 62% protein sequence identity to mouse Lipocalin-2.

  • 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 Lipocalin-2 Capture Antibody 1 x 600µl
    10X Human Lipocalin-2 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BR 1 x 6ml
    Human Lipocalin-2 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

    Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. Binds iron through association with 2,5-dihydroxybenzoic acid (2,5-DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. Iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Involved in apoptosis due to interleukin-3 (IL3) deprivation: iron-loaded form increases intracellular iron concentration without promoting apoptosis, while iron-free form decreases intracellular iron levels, inducing expression of the proapoptotic protein BCL2L11/BIM, resulting in apoptosis. Involved in innate immunity, possibly by sequestrating iron, leading to limit bacterial growth.
  • Tissue specificity

    Expressed in bone marrow and in tissues that are prone to exposure to microorganism. High expression is found in bone marrow as well as in uterus, prostate, salivary gland, stomach, appendix, colon, trachea and lung. Not found in the small intestine or peripheral blood leukocytes.
  • Sequence similarities

    Belongs to the calycin superfamily. Lipocalin family.
  • Cellular localization

    Secreted. Upon binding to the SLC22A17 (24p3R) receptor, it is internalized.
  • Information by UniProt
  • Alternative names

    • 24p3
    • 25 kDa alpha 2 microglobulin related subunit of MMP9
    • 25 kDa alpha-2-microglobulin-related subunit of MMP-9
    • Alpha 2 microglobulin related protein
    • HGNC:6526
    • HNL
    • Lcn 2
    • Lcn2
    • Lipocalin-2
    • Migration stimulating factor inhibitor
    • MSFI
    • Neutrophil gelatinase associated lipocalin
    • Neutrophil gelatinase associated lipocalin precursor
    • Neutrophil gelatinase-associated lipocalin
    • NGAL
    • NGAL_HUMAN
    • Oncogene 24p3
    • Oncogenic lipocalin 24p3
    • p25
    • Siderocalin
    • siderocalin LCN2
    • SV40 induced 24P3 protein
    • Uterocalin
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab215541 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 Lipocalin-2 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 2-fold increase in sensitivity.

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

  • The concentrations of Lipocalin-2 were measured in duplicates, interpolated from the Lipocalin-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 2%, plasma (citrate) 2%, plasma (EDTA) 2%, and plasma (heparin) 2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 61.4 ng/mL in serum, 52.2 ng/mL in plasma (citrate), 85.5ng/mL in plasma (EDTA) and 52.4ng/mL in plasma (heparin).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 77 ng/mL with a range of 37 – 135 ng/mL.

  • The concentrations of Lipocalin-2 were measured in duplicates, interpolated from the Lipocalin-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 3% and saliva 0.05%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 67.9 ng/mL in urine and 1,551ng/mL in saliva.

  • The concentrations of Lipocalin-2 were measured in duplicates, interpolated from the Lipocalin-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: A431 2% and PBMC 33%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 65.8 ng/mL in A431 and 4.85 ng/mL in PBMC cell culture supernatant.

Protocols

References

ab215541 has not yet been referenced specifically in any publications.

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