Human IL-12 (p70) PicoKine® Quick ELISA Kit

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.[i] 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.[ii] 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.[iii] The standard product used in this kit is recombinant human IL-12 p70 with the molecular mass of 75 KDa.

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 18 18 20 17 18 1.08 6% Sample 2 51 59 60 54 56 3.67 6.5% Sample 3 294 290 284 296 291 4.58 1.5% 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 18 18 20 17 18 1.08 6% Sample 1 18 18 20 17 18 1.08 6% Sample 2 51 59 60 54 56 3.67 6.5% Sample 2 51 59 60 54 56 3.67 6.5% Sample 3 294 290 284 296 291 4.58 1.5% Sample 3 294 290 284 296 291 4.58 1.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 18 51 294 19 55 310 Standard deviation 1.31 3.41 17.38 1.67 3.79 21.39 CV (%) 7.3% 6.7% 5.2% 8.8% 6.9% 6.9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 18 51 294 19 55 310 Standard deviation 1.31 3.41 17.38 1.67 3.79 21.39 CV (%) 7.3% 6.7% 5.2% 8.8% 6.9% 6.9% 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) 18 51 294 19 55 310 Mean (pg/ml) 18 51 294 19 55 310 Standard deviation 1.31 3.41 17.38 1.67 3.79 21.39 Standard deviation 1.31 3.41 17.38 1.67 3.79 21.39 CV (%) 7.3% 6.7% 5.2% 8.8% 6.9% 6.9% CV (%) 7.3% 6.7% 5.2% 8.8% 6.9% 6.9%

Description|Quantity|Volume Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 HRP-Linked Antibody|1|6ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.021 0.088 0.140 0.274 0.513 0.953 1.620 2.044 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.021 0.088 0.140 0.274 0.513 0.953 1.620 2.044 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 Concentration (pg/ml) 0 7.8 15.6 31.3 62.5 125 250 500 O.D. 0.021 0.088 0.140 0.274 0.513 0.953 1.620 2.044 O.D. 0.021 0.088 0.140 0.274 0.513 0.953 1.620 2.044

Microplate Reader capable of reading absorbance at 450nm. 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. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.