Rat 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 rat NGAL, consisting of 177 amino acids with the molecular mass of 21kDa.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 121 118 102 117 114 7.36 6.4% Sample 2 750 751 693 740 733 23.77 3.2% Sample 3 3188 2684 2847 3144 2965 208.95 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 1 121 118 102 117 114 7.36 6.4% Sample 1 121 118 102 117 114 7.36 6.4% Sample 2 750 751 693 740 733 23.77 3.2% Sample 2 750 751 693 740 733 23.77 3.2% Sample 3 3188 2684 2847 3144 2965 208.95 7% Sample 3 3188 2684 2847 3144 2965 208.95 7%

Neutrophil gelatinase-associated lipocalin; NGAL; Alpha-2-microglobulin-related protein; Alpha-2U globulin-related protein; Lipocalin-2; Siderocalin LCN2; p25; Lcn2

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 121 750 3188 126 727 3167 Standard deviation 6.05 50.25 168.96 7.3 56.7 177.35 CV (%) 5% 6.7% 5.3% 5.8% 7.8% 5.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 121 750 3188 126 727 3167 Standard deviation 6.05 50.25 168.96 7.3 56.7 177.35 CV (%) 5% 6.7% 5.3% 5.8% 7.8% 5.6% 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) 121 750 3188 126 727 3167 Mean (pg/ml) 121 750 3188 126 727 3167 Standard deviation 6.05 50.25 168.96 7.3 56.7 177.35 Standard deviation 6.05 50.25 168.96 7.3 56.7 177.35 CV (%) 5% 6.7% 5.3% 5.8% 7.8% 5.6% CV (%) 5% 6.7% 5.3% 5.8% 7.8% 5.6%

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 (By similarity) . .

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 78 156 312 625 1250 2500 5000 O.D. 0.031 0.145 0.225 0.418 0.739 1.09 1.661 2.13 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.031 0.145 0.225 0.418 0.739 1.09 1.661 2.13 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.031 0.145 0.225 0.418 0.739 1.09 1.661 2.13 O.D. 0.031 0.145 0.225 0.418 0.739 1.09 1.661 2.13

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.