Human Trail ELISA Kit EZ-SetTM (DIY Antibody Pairs)

In the field of cell biology, TNF-related apoptosis-inducing ligand (TRAIL), is a protein which functions as a ligand which induces the process of cell death called apoptosis. TRAIL has also been designated CD253 (cluster of differentiation 253) .Tumor necrosis factor (TNF) family cytokines function as prominent mediators of immune regulation and the inflammatory response. Most TNF family cytokines are expressed as type II transmembrane proteins, with homology confined to approximately 150 C-terminal residues. The TNF ligands interact with a parallel family of receptors. TRAIL binds to the death receptors DR4 (TRAIL-RI) and DR5 (TRAIL-RII) . The process of apoptosis is caspase-8-dependent. Caspase-8 activates downstream effector caspases including procaspase-3, -6, and -7, leading to activation of specific kinases. TRAIL also binds the receptors DcR1 and DcR2, which do not contain a cytoplasmic domain (DcR1) or contain a truncated death domain (DcR2) . The standard product used in this kit is recombinant human TRAIL with the molecular mass of 21KDa. It is expressed from the amino acids ectoderm T95--G28.

Tumor necrosis factor ligand superfamily member 10; Apo-2 ligand; Apo-2L; TNF-related apoptosis-inducing ligand; Protein TRAIL; CD253; TNFSF10; APO2L; TRAIL

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 38 184 416 34 175 377 Standard deviation 2.66 9.56 18.72 2.75 9.62 18.85 CV (%) 7% 5.2% 4.5% 8.1% 5.5% 5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 38 184 416 34 175 377 Standard deviation 2.66 9.56 18.72 2.75 9.62 18.85 CV (%) 7% 5.2% 4.5% 8.1% 5.5% 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) 38 184 416 34 175 377 Mean (pg/ml) 38 184 416 34 175 377 Standard deviation 2.66 9.56 18.72 2.75 9.62 18.85 Standard deviation 2.66 9.56 18.72 2.75 9.62 18.85 CV (%) 7% 5.2% 4.5% 8.1% 5.5% 5% CV (%) 7% 5.2% 4.5% 8.1% 5.5% 5%

Cytokine that binds to TNFRSF10A/TRAILR1, TNFRSF10B/TRAILR2, TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and possibly also to TNFRSF11B/OPG. Induces apoptosis. Its activity may be modulated by binding to the decoy receptors TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and TNFRSF11B/OPG that cannot induce apoptosis.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0532-CA Rabbit anti- human TNFSF10 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. EZ0532-DA Biotinylated goat anti- human TNFSF10 polyclonal antibody (Detection Antibody) 500 μl, 10 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0532-ST Lyophilized recombinant human TNFSF10 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 EZ0532-CA Rabbit anti- human TNFSF10 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. EZ0532-DA Biotinylated goat anti- human TNFSF10 polyclonal antibody (Detection Antibody) 500 μl, 10 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0532-ST Lyophilized recombinant human TNFSF10 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 EZ0532-CA Rabbit anti- human TNFSF10 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. EZ0532-CA Rabbit anti- human TNFSF10 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. EZ0532-DA Biotinylated goat anti- human TNFSF10 polyclonal antibody (Detection Antibody) 500 μl, 10 μg/mL (recommended dilution 1:100) EZ0532-DA Biotinylated goat anti- human TNFSF10 polyclonal antibody (Detection Antibody) 500 μl, 10 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) EK0532-ST Lyophilized recombinant human TNFSF10 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. EK0532-ST Lyophilized recombinant human TNFSF10 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 15.6 31.2 62.5 125 250 500 1000 O.D. 0.066 0.123 0.166 0.303 0.509 0.824 1.549 2.458 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.066 0.123 0.166 0.303 0.509 0.824 1.549 2.458 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.066 0.123 0.166 0.303 0.509 0.824 1.549 2.458 O.D. 0.066 0.123 0.166 0.303 0.509 0.824 1.549 2.458

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)