Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.313 pg/ml
- Range: 0.59 pg/ml - 2400 pg/ml
- Sample type: Cit plasma, Plasma, Serum, Urine
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Mouse
Overview
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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, Cit 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% Cit 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: Goat, Cow, 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.
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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.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 100X Stoplight Red Substrate 1 x 120µl 10X Mouse Lipocalin-2 Capture Antibody 1 x 600µl 10X Mouse Lipocalin-2 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl Antibody Diluent 5B 1 x 6ml Mouse Lipocalin-2 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Stoplight Red Substrate Buffer 1 x 12ml -
Research areas
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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
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Alternative names
- 24p3
- 25 kDa alpha 2 microglobulin related subunit of MMP9
- 25 kDa alpha-2-microglobulin-related subunit of MMP-9
see all -
Database links
- Entrez Gene: 16819 Mouse
- SwissProt: P11672 Mouse
- Unigene: 9537 Mouse
Images
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Other - Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)
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.
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Example of mouse Lipocalin-2 (NGAL) standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of Lipocalin-2 in mouse serum, plasma (citrate), and platelet poor plasma (EDTA).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).
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Comparison of secreted Lipocalin-2 in media, unstimulated and PMA/PHA-stimulated J774A.1 cells.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).
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Comparison of secreted Lipocalin-2 in mouse cell lines with or without stimulation.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.
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Sandwich ELISA - Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)To learn more about the advantages of recombinant antibodies see here.
