Human CD40/TNFRSF5 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

CD40 is a cell surface receptor that is expressed on the surface of all mature B cells, most mature B-cell malignancies, and some early B-cell acute lymphocytic leukemias, but is not expressed on plasma cells. The CD40 gene is localized to the long arm of chromosome 20, bands q12-q13.2. The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation.

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 93 325 538 101 357 529 Standard deviation 4.8 19.2 28.5 6.9 26.4 32.8 CV (%) 5.2% 5.9% 5.3% 6.8% 7.4% 6.2% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 93 325 538 101 357 529 Standard deviation 4.8 19.2 28.5 6.9 26.4 32.8 CV (%) 5.2% 5.9% 5.3% 6.8% 7.4% 6.2% 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) 93 325 538 101 357 529 Mean (pg/ml) 93 325 538 101 357 529 Standard deviation 4.8 19.2 28.5 6.9 26.4 32.8 Standard deviation 4.8 19.2 28.5 6.9 26.4 32.8 CV (%) 5.2% 5.9% 5.3% 6.8% 7.4% 6.2% CV (%) 5.2% 5.9% 5.3% 6.8% 7.4% 6.2%

Receptor for TNFSF5/CD40LG. Transduces TRAF6- and MAP3K8-mediated signals that activate ERK in macrophages and B cells, leading to induction of immunoglobulin secretion.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0702-CA Rabbit anti- human CD40 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. EZ0702-DA Biotinylated goat anti- human CD40 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0702-ST Lyophilized recombinant human CD40 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 EZ0702-CA Rabbit anti- human CD40 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. EZ0702-DA Biotinylated goat anti- human CD40 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0702-ST Lyophilized recombinant human CD40 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 EZ0702-CA Rabbit anti- human CD40 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. EZ0702-CA Rabbit anti- human CD40 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. EZ0702-DA Biotinylated goat anti- human CD40 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) EZ0702-DA Biotinylated goat anti- human CD40 polyclonal antibody (Detection Antibody) 500 μl, 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) EK0702-ST Lyophilized recombinant human CD40 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. EK0702-ST Lyophilized recombinant human CD40 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. 0.01 0.077 0.147 0.221 0.419 0.997 1.62 2.394 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.01 0.077 0.147 0.221 0.419 0.997 1.62 2.394 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. 0.01 0.077 0.147 0.221 0.419 0.997 1.62 2.394 O.D. 0.01 0.077 0.147 0.221 0.419 0.997 1.62 2.394

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)