Mouse Lipocalin-2 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Mouse Lipocalin-2 with a sensitivity of 0.313 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
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Key facts
- Detection method
- Fluorescent
- Sample types
- Urine, Plasma, Citrate plasma, Serum
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse
- Range
- 0.59 - 2400 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 0.313 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development (PubMed:12453413). Binds iron through association with 2,3-dihydroxybenzoic acid (2,3-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; limits bacterial proliferation by sequestering iron bound to microbial siderophores, such as enterobactin (PubMed:15531878, PubMed:16446425). Can also bind siderophores from M.tuberculosis (By similarity).
Alternative names
Neutrophil gelatinase-associated lipocalin, NGAL, Lipocalin-2, Oncogene 24p3, SV-40-induced 24p3 protein, Siderocalin LCN2, p25, 24p3, Lcn2
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Mouse Lipocalin-2 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Mouse Lipocalin-2 with a sensitivity of 0.313 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Urine, Plasma, Citrate plasma, Serum
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse
- Range
- 0.59 - 2400 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 0.313 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 5 | C.V. 3.7 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 3 | C.V. 6.1 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Serum | Average % = 109 | Range 106 - 110 % |
Sample type Plasma | Average % = 107 | Range 87 - 120 % |
Sample type Urine | Average % = 105 | Range 100 - 107 % |
Sample type Citrate plasma | Average % = 109 | Range 98 - 116 % |
Sample type Cell culture media | Average % = 104 | Range 102 - 107 % |
Sample type serum free media | Average % = 99 | Range 98 - 103 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
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.
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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4 product images
Sandwich ELISA - Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)
Example of mouse Lipocalin-2 (NGAL) standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)
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).
Sandwich ELISA - Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)
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).
Sandwich ELISA - Mouse Lipocalin-2 ELISA Kit, Fluorescent (ab229420)
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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