Overview

  • Product name
    Mouse Lipocalin-2 ELISA Kit, Fluorescent
    See all Lipocalin-2 / NGAL kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 5 3.7%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 6.1%
  • Sample type
    Urine, Serum, Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    0.313 pg/ml
  • Range
    0.59 pg/ml - 2400 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 105 100% - 107%
    Serum 109 106% - 110%
    Plasma 107 87% - 120%
    Cell culture media 104 102% - 107%
    Citrate Plasma 109 98% - 116%
    serum free media 99 98% - 103%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse
    Does not react with: Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Human, Pig
  • Product overview

    Lipocalin-2 (NGAL) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Lipocalin-2 (NGAL) protein in mouse serum, plasma, urine, and cell culture supernatant.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.


     

  • 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.  Mouse Lipocalin-2 has 81% and 62% protein sequence identity to rat and human Lipocalin-2, respectively.

  • 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 Mouse Lipocalin-2 Capture Antibody 1 x 600µl
    10X Mouse Lipocalin-2 Detector Antibody 1 x 600µl
    Mouse Lipocalin-2 Lyophilized Recombinant Protein 2 vials
    Antibody Diluent 5B 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Stoplight Red Substrate Buffer 1 x 12ml
    100X Stoplight Red Substrate 1 x 120µl
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Plate Seals 1 unit
  • 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 ab229420 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

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

  • The concentrations of Lipocalin-2 were measured in duplicate and interpolated from the Lipocalin-2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 265.2 ng/mL in mouse serum, 583.8 ng/mL in mouse plasma (Citrate), and 535.2 ng/mL in platelet poor mouse plasma (EDTA).

  • J774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. Raw data from duplicate measurements are plotted (mean +/- SD, n=2).

  • L929 and J774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. RAW264.7 were grown in the absence (unstimulated) or presence of LPS (stimulated) for 2 days. Cell Culture supernatant was diluted to within the range of the assay, 5-fold (L929 and J774A.1) or 2,000-fold (RAW264.7). Measured values were interpolated from the Lipocalin-2 Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed. Lipocalin-2 was undetectable in media.

Protocols

References

ab229420 has not yet been referenced specifically in any publications.

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