Human Elafin/PI3 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Elafin, also known as peptidase inhibitor 3 or skin-derived antileukoproteinase (SKALP) is a protein that in humans is encoded by the PI3 gene. It is mapped to 20q13.12. This gene encodes an elastase-specific inhibitor that functions as an antimicrobial peptide against Gram-positive and Gram-negative bacteria. The protein contains a WAP-type four-disulfide core (WFDC) domain, and is thus a member of the WFDC domain family. Most WFDC gene members are localized to chromosome 20q12-q13 in two clusters: centromeric and telomeric. This gene belongs to the centromeric cluster. Expression of this gene is upgulated by bacterial lipopolysaccharides and cytokines. This gene is the neutrophil and pancreatic elastase-specific inhibitor of skin. It may prevent elastase-mediated tissue proteolysis.

Elafin; Elastase-specific inhibitor; ESI; Peptidase inhibitor 3; PI-3; Protease inhibitor WAP3; Skin-derived antileukoproteinase; SKALP; WAP four-disulfide core domain protein 14; PI3; WAP3, WFDC14

Cell Biology, Cell Cycle, Immunology, Innate Immunity, Kinases/Phosphatases, Lipid Signaling, Protein Phosphorylation, Ser/Thr Kinases, Signal Transduction, Signaling Pathway, TLR Signaling

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 225 657 1334 230 645 1742 Standard deviation 12.6 32.2 57.36 17.02 39.99 113.23 CV (%) 5.6% 4.9% 4.3% 7.4% 6.2% 6.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 225 657 1334 230 645 1742 Standard deviation 12.6 32.2 57.36 17.02 39.99 113.23 CV (%) 5.6% 4.9% 4.3% 7.4% 6.2% 6.5% 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) 225 657 1334 230 645 1742 Mean (pg/ml) 225 657 1334 230 645 1742 Standard deviation 12.6 32.2 57.36 17.02 39.99 113.23 Standard deviation 12.6 32.2 57.36 17.02 39.99 113.23 CV (%) 5.6% 4.9% 4.3% 7.4% 6.2% 6.5% CV (%) 5.6% 4.9% 4.3% 7.4% 6.2% 6.5%

Catalog number Description Quantity Storage of opened/reconstituted material EZ1117-CA Rabbit anti- human PI3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1117-DA Biotinylated goat anti- human PI3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1117-ST Lyophilized recombinant human PI3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1117-CA Rabbit anti- human PI3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1117-DA Biotinylated goat anti- human PI3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1117-ST Lyophilized recombinant human PI3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1117-CA Rabbit anti- human PI3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1117-CA Rabbit anti- human PI3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1117-DA Biotinylated goat anti- human PI3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) EZ1117-DA Biotinylated goat anti- human PI3 polyclonal antibody (Detection Antibody) 500 μl, 3.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1117-ST Lyophilized recombinant human PI3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK1117-ST Lyophilized recombinant human PI3 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

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 31.2 62.5 125 250 500 1000 2000 O.D. 0.01 0.073 0.148 0.271 0.575 0.966 1.789 2.233 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.01 0.073 0.148 0.271 0.575 0.966 1.789 2.233 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.01 0.073 0.148 0.271 0.575 0.966 1.789 2.233 O.D. 0.01 0.073 0.148 0.271 0.575 0.966 1.789 2.233

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)