LCN2 Chemi-Luminescent ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Neutrophil gelatinase-associated lipocalin (NGAL) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Neutrophil gelatinase-associated lipocalin (NGAL) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

24p3;25 kDa alpha-2-microglobulin-related subunit of MMP-9; Lipocalin-2; migration-stimulating factor inhibitor; MSFI; neutrophil gelatinase-associated lipocalin; NGAL; oncogene 24p3; p25; siderocalin 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.

The microplate provided in this kit has been pre-coated with an antibody specific to Neutrophil gelatinase-associated lipocalin (NGAL) . Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Neutrophil gelatinase-associated lipocalin (NGAL) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Neutrophil gelatinase-associated lipocalin (NGAL) level in the sample or standard.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 82-105 101 EDTA Plasma (n=5) 90-101 94 Heparin Plasma (n=5) 87-101 92 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 82-105 101 Serum (n=5) 82-105 101 EDTA Plasma (n=5) 90-101 94 EDTA Plasma (n=5) 90-101 94 Heparin Plasma (n=5) 87-101 92 Heparin Plasma (n=5) 87-101 92

Component Amount LCN2 Microplate 96 Wells (12 x 8 Well strips) LCN2 Lyophilized Standard 2 100X LCN2 Biotinylated Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 100X Luminol Substrate 1 x 2 mL Substrate Diluent 1 x 20 mL Component Amount Component Amount LCN2 Microplate 96 Wells (12 x 8 Well strips) LCN2 Microplate 96 Wells (12 x 8 Well strips) LCN2 Lyophilized Standard 2 LCN2 Lyophilized Standard 2 100X LCN2 Biotinylated Detector Anitbody 1 x 120 uL 100X LCN2 Biotinylated Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL 100X Luminol Substrate 1 x 2 mL 100X Luminol Substrate 1 x 2 mL Substrate Diluent 1 x 20 mL Substrate Diluent 1 x 20 mL