Rat HE4/WFDC2 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

WAP four-disulfide core domain protein 2, also known as Human Epididymis Protein 4 (HE4), is a protein that in humans is encoded by the WFDC2 gene. This gene encodes a protein that is a member of the WFDC domain family. The WFDC domain, or WAP Signature motif, contains eight cysteines forming four disulfide bonds at the core of the protein, and functions as a protease inhibitor in many family members. This gene is expressed in pulmonary epithelial cells, and was also found to be expressed in some ovarian cancers. The encoded protein is a small secretory protein, which may be involved in sperm maturation.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 422 1434 8876 525 1688 8921 Standard deviation 16.04 61.66 470.43 22.05 82.71 517.42 CV (%) 3.8% 4.3% 5.3% 4.2% 4.9% 5.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 422 1434 8876 525 1688 8921 Standard deviation 16.04 61.66 470.43 22.05 82.71 517.42 CV (%) 3.8% 4.3% 5.3% 4.2% 4.9% 5.8% 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) 422 1434 8876 525 1688 8921 Mean (pg/ml) 422 1434 8876 525 1688 8921 Standard deviation 16.04 61.66 470.43 22.05 82.71 517.42 Standard deviation 16.04 61.66 470.43 22.05 82.71 517.42 CV (%) 3.8% 4.3% 5.3% 4.2% 4.9% 5.8% CV (%) 3.8% 4.3% 5.3% 4.2% 4.9% 5.8%

Catalog number Description Quantity Storage of opened/reconstituted material EZ1624-CA Rabbit anti- rat Wfdc2 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. EZ1624-DA Biotinylated goat anti- rat Wfdc2 polyclonal antibody (Detection Antibody) 500 μl, 6.25 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1624-ST Lyophilized recombinant rat Wfdc2 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 EZ1624-CA Rabbit anti- rat Wfdc2 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. EZ1624-DA Biotinylated goat anti- rat Wfdc2 polyclonal antibody (Detection Antibody) 500 μl, 6.25 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1624-ST Lyophilized recombinant rat Wfdc2 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 EZ1624-CA Rabbit anti- rat Wfdc2 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. EZ1624-CA Rabbit anti- rat Wfdc2 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. EZ1624-DA Biotinylated goat anti- rat Wfdc2 polyclonal antibody (Detection Antibody) 500 μl, 6.25 μg/mL (recommended dilution 1:100) EZ1624-DA Biotinylated goat anti- rat Wfdc2 polyclonal antibody (Detection Antibody) 500 μl, 6.25 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) EK1624-ST Lyophilized recombinant rat Wfdc2 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. EK1624-ST Lyophilized recombinant rat Wfdc2 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) 0 156 312 625 1250 2500 5000 10000 O.D. 0.082 0.173 0.225 0.311 0.496 0.819 1.297 2.064 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.082 0.173 0.225 0.311 0.496 0.819 1.297 2.064 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.082 0.173 0.225 0.311 0.496 0.819 1.297 2.064 O.D. 0.082 0.173 0.225 0.311 0.496 0.819 1.297 2.064

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)