Mouse NGAL (Neutrophil Gelatinase Associated Lipocalin) ELISA Kit (MOES01298)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- LCN2, Lipocalin 2, Oncogene 24p3, MSFI
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||39.06-2500 pg/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Mouse NGAL in samples. No significant cross-reactivity or interference between Mouse NGAL and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse NGAL. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse NGAL and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse NGAL, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse NGAL. The concentration of Mouse NGAL in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||LCN2: 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. Belongs to the calycin superfamily. Lipocalin family. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Secreted; Secreted, signal peptide
Cellular Component: extracellular space; cell; extracellular region; cytosol
Molecular Function:protein binding; protein homodimerization activity; protease binding; transporter activity; iron ion binding
Biological Process: response to drug; iron ion homeostasis; immune system process; transport; apoptosis; response to virus; siderophore transport; positive regulation of cell projection organization and biogenesis; innate immune response; ion transport; response to herbicide
|NCBI GenInfo Identifier:||112725|
|NCBI Gene ID:||16819|
|NCBI Accession:||P11672. 1|
|UniProt Secondary Accession:||P11672,Q3UE34,|
|UniProt Related Accession:||P11672|
|Molecular Weight:||22,875 Da|
|NCBI Full Name:||Neutrophil gelatinase-associated lipocalin|
|NCBI Synonym Full Names:||lipocalin 2|
|NCBI Official Symbol:||Lcn2|
|NCBI Official Synonym Symbols:||24p3; Sip24; AW212229|
|NCBI Protein Information:||neutrophil gelatinase-associated lipocalin; p25; lipocalin-2; siderocalin LCN2; SV-40-induced 24P3 protein; secreted inducible protein 24|
|UniProt Protein Name:||Neutrophil gelatinase-associated lipocalin|
|UniProt Synonym Protein Names:||Lipocalin-2; SV-40-induced 24P3 protein; Siderocalin LCN2; p25|
|Protein Family:||Neutrophil gelatinase-associated lipocalin|
|UniProt Gene Name:||Lcn2|
|UniProt Entry Name:||NGAL_MOUSE|
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse NGAL were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse NGAL were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.80||4.43||3.99||6.57||5.34||5.34|
The recovery of Mouse NGAL spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||89-102||95|
|Cell culture media (n=5)||91-108||99|
Samples were spiked with high concentrations of Mouse NGAL and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20°C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 µL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 µL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.