Human Lipocalin-2/NGAL ELISA Kit PicoKine®

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.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 256 230 223 260 242 16 6.6% Sample 2 1510 1658 1546 1807 1630 115.72 7% Sample 3 5422 5479 5362 4730 5248 302.05 5.7% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 256 230 223 260 242 16 6.6% Sample 1 256 230 223 260 242 16 6.6% Sample 2 1510 1658 1546 1807 1630 115.72 7% Sample 2 1510 1658 1546 1807 1630 115.72 7% Sample 3 5422 5479 5362 4730 5248 302.05 5.7% Sample 3 5422 5479 5362 4730 5248 302.05 5.7%

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. .

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 256 1510 5422 254 1537 5138 Standard deviation 19.2 81.54 281.94 19.05 84.53 323.69 CV (%) 7.5% 5.4% 5.2% 7.5% 5.5% 6.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 256 1510 5422 254 1537 5138 Standard deviation 19.2 81.54 281.94 19.05 84.53 323.69 CV (%) 7.5% 5.4% 5.2% 7.5% 5.5% 6.3% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 256 1510 5422 254 1537 5138 Mean (pg/ml) 256 1510 5422 254 1537 5138 Standard deviation 19.2 81.54 281.94 19.05 84.53 323.69 Standard deviation 19.2 81.54 281.94 19.05 84.53 323.69 CV (%) 7.5% 5.4% 5.2% 7.5% 5.5% 6.3% CV (%) 7.5% 5.4% 5.2% 7.5% 5.5% 6.3%

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. .

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.007 0.084 0.174 0.308 0.585 1.076 1.516 1.99 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.007 0.084 0.174 0.308 0.585 1.076 1.516 1.99 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.007 0.084 0.174 0.308 0.585 1.076 1.516 1.99 O.D. 0.007 0.084 0.174 0.308 0.585 1.076 1.516 1.99

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.