Mouse IL-12 (p70) ELISA Kit PicoKine®

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.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 37 38 39 40 38 1.11 2.9% Sample 2 150 151 171 168 160 9.56 5.9% Sample 3 410 416 442 417 421 12.27 2.9% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 37 38 39 40 38 1.11 2.9% Sample 1 37 38 39 40 38 1.11 2.9% Sample 2 150 151 171 168 160 9.56 5.9% Sample 2 150 151 171 168 160 9.56 5.9% Sample 3 410 416 442 417 421 12.27 2.9% Sample 3 410 416 442 417 421 12.27 2.9%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 37 150 410 33 147 442 Standard deviation 2.18 9.15 30.75 2.44 8.96 38.45 CV (%) 7.5% 6.1% 7.5% 7.4% 6.1% 8.7% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 37 150 410 33 147 442 Standard deviation 2.18 9.15 30.75 2.44 8.96 38.45 CV (%) 7.5% 6.1% 7.5% 7.4% 6.1% 8.7% 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) 37 150 410 33 147 442 Mean (pg/ml) 37 150 410 33 147 442 Standard deviation 2.18 9.15 30.75 2.44 8.96 38.45 Standard deviation 2.18 9.15 30.75 2.44 8.96 38.45 CV (%) 7.5% 6.1% 7.5% 7.4% 6.1% 8.7% CV (%) 7.5% 6.1% 7.5% 7.4% 6.1% 8.7%

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

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. 0.003 0.056 0.097 0.203 0.388 0.691 1.286 1.906 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.003 0.056 0.097 0.203 0.388 0.691 1.286 1.906 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. 0.003 0.056 0.097 0.203 0.388 0.691 1.286 1.906 O.D. 0.003 0.056 0.097 0.203 0.388 0.691 1.286 1.906

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.