| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-12 subunit alpha; IL-12A; Cytotoxic lymphocyte maturation factor 35 kDa subunit; CLMF p35; IL-12 subunit p35; NK cell stimulatory factor chain 1; NKSF1; IL12A |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: (I23-S328) + (R23-S219) |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human interleukin-12 subunit alpha (gene IL12A) in cell culture supernatants and serum. 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.
IL12A encodes the p35 subunit, which pairs with p40 (IL12B) to form interleukin-12 (IL-12 p70) or with EBI3 to form IL-35. IL-12 is produced by dendritic cells and macrophages, drives IFN-γ production by T and NK cells and promotes Th1 differentiation, so it is a key link between innate and adaptive immunity.
Kit Components
| Anti-Human IL12A Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human IL12A Standard | 2 vials, 10 ng/tube |
| Human IL12A 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 | Human |
|---|---|
| Sample types | Cell culture supernatants and serum |
| Assay range | 7.8 pg/mL–500 pg/mL |
| Sensitivity | <2 pg/mL |
| Standard | Recombinant human IL12A ((I23-S328)), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.2–7.3% (3 samples, n = 16 each) |
| Inter-assay CV | 6.9–8.8% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum samples measured 2–50 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.003; 7.8 = 0.056; 15.6 = 0.097; 31.3 = 0.203; 62.5 = 0.388; 125 = 0.691; 250 = 1.286; 500 = 1.906. 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 | 7.8 | 15.6 | 31.3 | 62.5 | 125 | 250 | 500 |
| O.D. | 0.003 | 0.056 | 0.097 | 0.203 | 0.388 | 0.691 | 1.286 | 1.906 |
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) | 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% |
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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