Rat Lipocalin-2/NGAL PicoKine® Quick ELISA Kit

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 1 117 122 105 113 114 7.20 6.3% Sample 2 762 763 685 730 735 27.20 3.7% Sample 3 3264 2569 2963 3288 3021 199.39 6.6% 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 1 117 122 105 113 114 7.20 6.3% Sample 1 117 122 105 113 114 7.20 6.3% Sample 2 762 763 685 730 735 27.20 3.7% Sample 2 762 763 685 730 735 27.20 3.7% Sample 3 3264 2569 2963 3288 3021 199.39 6.6% Sample 3 3264 2569 2963 3288 3021 199.39 6.6%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 117 762 3264 133 782 3365 Standard deviation 6.08 49.53 172.99 7.58 60.21 185.08 CV (%) 5.2% 6.5% 5.3% 5.7% 7.7% 5.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 117 762 3264 133 782 3365 Standard deviation 6.08 49.53 172.99 7.58 60.21 185.08 CV (%) 5.2% 6.5% 5.3% 5.7% 7.7% 5.5% 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) 117 762 3264 133 782 3365 Mean (pg/ml) 117 762 3264 133 782 3365 Standard deviation 6.08 49.53 172.99 7.58 60.21 185.08 Standard deviation 6.08 49.53 172.99 7.58 60.21 185.08 CV (%) 5.2% 6.5% 5.3% 5.7% 7.7% 5.5% CV (%) 5.2% 6.5% 5.3% 5.7% 7.7% 5.5%

Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.020 0.061 0.128 0.236 0.412 0.835 1.390 2.107 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.020 0.061 0.128 0.236 0.412 0.835 1.390 2.107 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.020 0.061 0.128 0.236 0.412 0.835 1.390 2.107 O.D. 0.020 0.061 0.128 0.236 0.412 0.835 1.390 2.107

Microplate Reader capable of reading absorbance at 450nm. 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. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.