Mouse TNFRSF9/4-1BB ELISA Kit EZ-SetTM (DIY Antibody Pairs)

CD137 is a member of the tumor necrosis factor (TNF) receptor family. Its alternative names are tumor necrosis factor receptor superfamily member 9 (TNFRSF9), 4-1BB and induced by lymphocyte activation (ILA) . CD137 gene was localized to chromosome 1p36.Members of this receptor family and their structurally related ligands are important regulators of a wide variety of physiological processes and play an especially important role in the regulation of immune responses. CD137 can be expressed by activated T cells, but to a larger extent on CD8 than on CD4 T cells. The best characterized activity of CD137 is its costimulatory activity for activated T cells. Crosslinking of CD137 enhances T cell proliferation, IL-2 secretion survival and cytolytic activity. Further, it can enhance immune activity to eliminate tumors in mice.

Apoptosis, Associated Proteins, Cancer, Cell Biology, Cell Death, Cell Type Markers, Cytokines, Hematopoietic Progenitors, Immunology, Innate Immunity, Lymphoid, Receptors, Stem Cells, T Lymphocytic Lineage, TNF Superfamily, Tumor Immunology

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 319 746 1253 286 833 1353 Standard deviation 12.76 28.3 66.5 16.6 37.5 94.7 CV (%) 4% 3.8% 5.3% 5.8% 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) 319 746 1253 286 833 1353 Standard deviation 12.76 28.3 66.5 16.6 37.5 94.7 CV (%) 4% 3.8% 5.3% 5.8% 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) 319 746 1253 286 833 1353 Mean (pg/ml) 319 746 1253 286 833 1353 Standard deviation 12.76 28.3 66.5 16.6 37.5 94.7 Standard deviation 12.76 28.3 66.5 16.6 37.5 94.7 CV (%) 4% 3.8% 5.3% 5.8% 4.5% 7% CV (%) 4% 3.8% 5.3% 5.8% 4.5% 7%

Catalog number Description Quantity Storage of opened/reconstituted material EZ1143-CA Rabbit anti- mouse Tnfrsf9 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. EZ1143-DA Biotinylated goat anti- mouse Tnfrsf9 polyclonal antibody (Detection Antibody) 500 μl, 5.5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1143-ST Lyophilized recombinant mouse Tnfrsf9 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 EZ1143-CA Rabbit anti- mouse Tnfrsf9 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. EZ1143-DA Biotinylated goat anti- mouse Tnfrsf9 polyclonal antibody (Detection Antibody) 500 μl, 5.5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1143-ST Lyophilized recombinant mouse Tnfrsf9 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 EZ1143-CA Rabbit anti- mouse Tnfrsf9 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. EZ1143-CA Rabbit anti- mouse Tnfrsf9 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. EZ1143-DA Biotinylated goat anti- mouse Tnfrsf9 polyclonal antibody (Detection Antibody) 500 μl, 5.5 μg/mL (recommended dilution 1:100) EZ1143-DA Biotinylated goat anti- mouse Tnfrsf9 polyclonal antibody (Detection Antibody) 500 μl, 5.5 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) EK1143-ST Lyophilized recombinant mouse Tnfrsf9 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. EK1143-ST Lyophilized recombinant mouse Tnfrsf9 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. 0.039 0.066 0.122 0.206 0.372 0.748 1.375 2.121 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.039 0.066 0.122 0.206 0.372 0.748 1.375 2.121 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. 0.039 0.066 0.122 0.206 0.372 0.748 1.375 2.121 O.D. 0.039 0.066 0.122 0.206 0.372 0.748 1.375 2.121

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)