| 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 one-step sandwich ELISA quantifies human interleukin-12 subunit alpha (gene IL12A) in cell culture supernatants and serum. The strips are pre-coated with a capture antibody; sample and an HRP-linked detection antibody are incubated together, so the assay needs fewer wash and incubation steps than a standard sandwich ELISA (the supplier markets the Quick format as a 90-minute assay). TMB develops color in proportion to the bound target, and the plate is read at 450 nm.
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 IL-12(p70) Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human IL-12(p70) Standard | 2 × 10 ng/tube |
| HRP Linked Anti-Human IL-12(p70) Antibody | 6 mL |
| Sample Diluent | 15 mL |
| TBS-T Wash Buffer (25x) | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Adhesive Plate Sealers | 2 |
Also required (not supplied): reader capable of reading absorbance at 450 nm; 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; horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3 mm.
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 |
Storage & Handling
Store at 4 °C for up to 6 months. Ships on gel ice and tolerates up to 3 days at room temperature; refrigerate on arrival.
Safety
Sample data
| 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 |
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|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|PieceMaterials required but not provided
- 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.
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