Human Neutrophil Elastase/ELA2/ELANE ELISA Kit PicoKine®

In humans, neutrophil elastase is encoded by the ELANE gene, which resides on chromosome 19. It is mapped to 19p13.3. Elastases form a subfamily of serine proteases that hydrolyze many proteins in addition to elastin. Humans have six elastase genes which encode structurally similar proteins. The encoded preproprotein is proteolytically processed to generate the active protease. Following activation, this protease hydrolyzes proteins within specialized neutrophil lysosomes, called azurophil granules, as well as proteins of the extracellular matrix. The enzyme may play a role in degenerative and inflammatory diseases through proteolysis of collagen-IV and elastin. This protein also degrades the outer membrane protein A (OmpA) of E. coli as well as the virulence factors of such bacteria as Shigella, Salmonella and Yersinia. Mutations in this gene are associated with cyclic neutropenia and severe congenital neutropenia (SCN) . This gene is present in a gene cluster on chromosome 19.

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 1174 1071 1108 1106 1114 37.23 3.3% Sample 2 5107 5193 4515 4791 4901 268.69 5.4% Sample 3 21947 21460 19663 20115 20796 937.16 4.5% 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 1174 1071 1108 1106 1114 37.23 3.3% Sample 1 1174 1071 1108 1106 1114 37.23 3.3% Sample 2 5107 5193 4515 4791 4901 268.69 5.4% Sample 2 5107 5193 4515 4791 4901 268.69 5.4% Sample 3 21947 21460 19663 20115 20796 937.16 4.5% Sample 3 21947 21460 19663 20115 20796 937.16 4.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1174 5107 21947 1365 6165 23984 Standard deviation 48.13 214.49 987.62 64.16 332.91 1367.09 CV (%) 4.1% 4.2% 4.5% 4.7% 5.4% 5.7% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1174 5107 21947 1365 6165 23984 Standard deviation 48.13 214.49 987.62 64.16 332.91 1367.09 CV (%) 4.1% 4.2% 4.5% 4.7% 5.4% 5.7% 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) 1174 5107 21947 1365 6165 23984 Mean (pg/ml) 1174 5107 21947 1365 6165 23984 Standard deviation 48.13 214.49 987.62 64.16 332.91 1367.09 Standard deviation 48.13 214.49 987.62 64.16 332.91 1367.09 CV (%) 4.1% 4.2% 4.5% 4.7% 5.4% 5.7% CV (%) 4.1% 4.2% 4.5% 4.7% 5.4% 5.7%

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 (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.057 0.091 0.124 0.241 0.418 0.87 1.736 2.447 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.057 0.091 0.124 0.241 0.418 0.87 1.736 2.447 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.057 0.091 0.124 0.241 0.418 0.87 1.736 2.447 O.D. 0.057 0.091 0.124 0.241 0.418 0.87 1.736 2.447

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.