Catalog No: OKBB00344
Size:96 Tests
Price: $466.00
SKU
OKBB00344
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Lipocalin-2/NGAL ELISA Kit (Human) (OKBB00344)

Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityHuman
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva's human NGAL ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for NGAL has been precoated onto 96-well plates. Standards(NS0, Q21-G198) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for NGAL is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human NGAL amount of sample captured in plate.
ELISA Kit Range156 pg/ml - 10,000 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)124634866827152235947133
St. Dev.48.6174.3423.3123.3230492.2
%CV3.9056.208.106.396.90
ELISA Kit Component
ComponentAmount
Lyophilized recombinant human NGAL standard10 ng/tube x 2
Anti-human NGAL Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-human NGAL antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Additional InformationRange: 156pg/ml-10, 000pg/ml
::Principle: Aviva’s human NGAL ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for NGAL has been precoated onto 96-well plates. Standards(NSO, Q21-G198) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for NGAL is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase
Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human NGAL amount of sample captured in plate.
::Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before using.
Background: Lipocalin-2 (LCN2), also known as NGAL, is a protein associated with neutrophil gelatinase.1 The LCN2 gene is located at 9q34 and contains 7 exons.2 The 25-kD LCN2 protein is believed to bind small lipophilic substances such as bacteria-derived lipopolysaccharide (LPS) and formylpeptides and may function as a modulator of inflammation. NGAL tightly binds bacterial catecholate-type ferric siderophores through a cyclically permuted, hybrid electrostatic/cation-pi interaction and is a potent bacteriostatic agent in iron-limiting conditions.3 The primary LCN2 transcript is 3,696 nucleotides long, and the processed transcript is 809 nucleotides long.4 LCN2 expression in adult bone marrow, uterus, prostate, salivary gland, stomach, appendix, colon, trachea, and lung, and in fetal spleen and lung. The standard product used in this kit is recombinant human NGAL, consisting of 178 amino acids with the molecular mass of 22KDa.
Reconstitution and StorageStore at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
Sample Typecell culture supernates, serum, plasma (heparin), saliva and urine
Sensitivity<10 pg/ml
Predicted Homology Based on Immunogen SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. Kjeldsen, L.; Johnsen, A. H.; Sengelov, H.; Borregaard, N. : Isolation and primary structure of NGAL, a novel protein
associated with human neutrophil gelatinase. J. Biol. Chem. 268: 10425-10432, 1993.
2. Cowland, J. B.; Borregaard, N. : Molecular characterization and pattern of tissue expression of the gene for
neutrophil gelatinase-associated lipocalin from humans. Genomics 45: 17-23, 1997.
3. Goetz, D. H.; Holmes, M. A.; Borregaard, N.; Bluhm, M. E.; Raymond, K. N.; Strong, R. K. : The neutrophil lipocalin
NGAL is a bacteriostatic agent that interferes with siderophore-mediated iron acquisition. Molec. Cell 10: 1033-1043,
2002.
4. Cowland, J. B.; Borregaard, N. : Molecular characterization and pattern of tissue expression of the gene for
neutrophil gelatinase-associated lipocalin from humans. Genomics 45: 17-23, 1997.
SpecificityNatural and recombinant human NGAL
ImmunogenExpression system for standard: NS0; Immunogen sequence: Q21-G198
DescriptionFor quantitative detection of human NGAL in cell culture supernatants, serum, plasma(heparin), saliva and urine.
Gene SymbolLCN2
Gene Full Namelipocalin 2
Alias Symbols24p3, 25 kDa alpha-2-microglobulin-related subunit of MMP-9, HNL, Lcn2, Lipocalin 2, Lipocalin-2, Migration stimulating factor inhibitor, MSFI, Neutrophil gelatinase-associated lipocalin, NGAL, NGAL_HUMAN, Oncogene 24p3, p25, Siderocalin, Siderocalin LCN2
NCBI Gene Id3934
Protein NameNeutrophil gelatinase-associated lipocalin
Description of TargetLipocalin-2 (LCN2), also known as NGAL, is a protein associated with neutrophil gelatinase.1 The LCN2 gene is located at 9q34 and contains 7 exons.2 The 25-kD LCN2 protein is believed to bind small lipophilic substances such as bacteria-derived lipopolysaccharide (LPS) and formylpeptides and may function as a modulator of inflammation. NGAL tightly binds bacterial catecholate-type ferric siderophores through a cyclically permuted, hybrid electrostatic/cation-pi interaction and is a potent bacteriostatic agent in iron-limiting conditions.3 The primary LCN2 transcript is 3,696 nucleotides long, and the processed transcript is 809 nucleotides long.4 LCN2 expression in adult bone marrow, uterus, prostate, salivary gland, stomach, appendix, colon, trachea, and lung, and in fetal spleen and lung. The standard product used in this kit is recombinant human NGAL, consisting of 178 amino acids with the molecular mass of 22KDa.
Uniprot IDP80188
Protein Accession #NP_005555.2
Nucleotide Accession #NM_005564.4
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