Mouse IL-12 (p70) ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Interleukin (IL) -12 is a 70-KDa cytokine comprised of two disulfide-linked proteins (p35 and p40) and is essential for the initiation of effective immune response. And the IL-12p70 is a heterodimer of p35 and p40 subunits; it is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation. Gene expression analysis of the IL-12 cytokine family subunits revealed that both strains induced high levels of p40 (protein chain communal to IL-12 p70 and IL-23) as well as p19, a subunit of IL-23. Conversely only ACT- 18HS19 infection induced consistent transcription of IL-12 p35, a subunit of IL-12 p70. The standard product used in this kit is recombinant mouse IL-12 p70 comprising of a 193 amino acids p35 chain and a 313 amino acids p40 chain connected by a disulfide bond.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0422-CA Mouse anti- mouse IL-12 (p70) monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0422-DA Biotinylated goat anti- mouse IL-12 (p70) polyclonal antibody (Detection Antibody) 500 μl, 0.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0422-ST Lyophilized recombinant Mouse anti- mouse IL-12 (p70) monoclonal 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 EZ0422-CA Mouse anti- mouse IL-12 (p70) monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0422-DA Biotinylated goat anti- mouse IL-12 (p70) polyclonal antibody (Detection Antibody) 500 μl, 0.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0422-ST Lyophilized recombinant Mouse anti- mouse IL-12 (p70) monoclonal 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 EZ0422-CA Mouse anti- mouse IL-12 (p70) monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0422-CA Mouse anti- mouse IL-12 (p70) monoclonal antibody (Capture Antibody) 500 μl, 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. EZ0422-DA Biotinylated goat anti- mouse IL-12 (p70) polyclonal antibody (Detection Antibody) 500 μl, 0.3 μg/mL (recommended dilution 1:100) EZ0422-DA Biotinylated goat anti- mouse IL-12 (p70) polyclonal antibody (Detection Antibody) 500 μl, 0.3 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) EK0422-ST Lyophilized recombinant Mouse anti- mouse IL-12 (p70) monoclonal 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. EK0422-ST Lyophilized recombinant Mouse anti- mouse IL-12 (p70) monoclonal 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. 0.041 0.101 0.159 0.270 0.459 0.869 1.507 2.092 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.041 0.101 0.159 0.270 0.459 0.869 1.507 2.092 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. 0.041 0.101 0.159 0.270 0.459 0.869 1.507 2.092 O.D. 0.041 0.101 0.159 0.270 0.459 0.869 1.507 2.092

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)