| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-12 subunit beta;IL-12B;Cytotoxic lymphocyte maturation factor 40 kDa subunit;CLMF p40;IL-12 subunit p40;Il12b; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: M23-S335 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies mouse interleukin-12 subunit beta (gene Il12b) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
IL12B encodes the p40 subunit shared by interleukin-12 (with p35) and interleukin-23 (with p19). IL-12 promotes Th1 responses and IFN-γ production, while IL-23 supports Th17 responses, which makes p40 central to studies of cellular immunity and inflammatory disease.
Kit Components
| Anti-Mouse Il12b Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Il12b Standard | 2 vials, 10 ng/tube |
| Mouse Il12b Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant mouse Il12b (Met23–Ser335), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.3–6.0% (3 samples, n = 16 each) |
| Inter-assay CV | 5.6–6.9% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum and plasma samples measured about 300 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.023; 15.6 = 0.093; 31.2 = 0.158; 62.5 = 0.271; 125 = 0.504; 250 = 0.878; 500 = 1.344; 1000 = 1.938. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.023 | 0.093 | 0.158 | 0.271 | 0.504 | 0.878 | 1.344 | 1.938 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 33 | 233 | 425 | 35 | 253 | 463 |
| Standard deviation | 1.84 | 13.98 | 22.52 | 1.96 | 17.45 | 29.63 |
| CV (%) | 6% | 6% | 5.3% | 5.6% | 6.9% | 6.4% |
Kit components
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|4Materials required but not provided
- 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.
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