| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-12 subunit alpha; IL-12A; Cytotoxic lymphocyte maturation factor 35 kDa subunit; CLMF p35; IL-12 subunit p35; Il12a |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: (M23 - S335) + (R23 - S215) |
| 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 rat 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-Rat Il12a Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Rat Il12a Standard | 2 vials, 10 ng/tube |
| Rat 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 | Rat |
|---|---|
| Sample types | Cell culture supernatants and serum |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Sensitivity | <1.5 pg/mL |
| Standard | Recombinant rat Il12a ((M23), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.5–7.1% (3 samples, n = 16 each) |
| Inter-assay CV | 6.1–8.9% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:10. In the supplier’s tests, serum samples measured 600–3200 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.044; 15.6 = 0.111; 31.2 = 0.17; 62.5 = 0.3; 125 = 0.558; 250 = 0.92; 500 = 1.518; 1000 = 2.216. 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. | 0.044 | 0.111 | 0.17 | 0.3 | 0.558 | 0.92 | 1.518 | 2.216 |
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) | 35 | 222 | 502 | 34 | 224 | 454 |
| Standard deviation | 1.92 | 15.76 | 27.61 | 2.07 | 19.93 | 29.05 |
| CV (%) | 5.5% | 7.1% | 5.5% | 6.1% | 8.9% | 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.
Can’t Find What You’re Looking For? We can help you source the best match or customize an ELISA solution for your study. Options may include alternative target synonyms, different species reactivity, sample type/matrix compatibility (serum/plasma/lysate/supernatant), assay format (sandwich/competitive), sensitivity/range, detection chemistry (colorimetric/fluorescent/chemiluminescent), plate format (pre-coated/uncoated, strips vs full plate), and bulk or custom packaging. Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
Shaoguang Li et al. (2023) Implantable Hydrogel-Protective DNA Aptamer-Based Sensor Supports Accurate, Continuous Electrochemical Analysis of Drugs at Multiple Sites in Living Rats. ACS Nano. 10.1021/acsnano.3c06520
Yue Li et al. (2022) Lactobacillus casei Improve Anti-Tuberculosis Drugs-Induced Intestinal Adverse Reactions in Rat by Modulating Gut Microbiota and Short-Chain Fatty Acids. Nutrients. 10.3390/nu14081668
Zhao Xin et al. (2019) Saikosaponin A Inhibits Breast Cancer by Regulating Th1/Th2 Balance. Frontiers in Pharmacology. 10.3389/fphar.2019.00624
Qian Zhou et al. (2019) Flaxseed oil alleviates dextran sulphate sodium-induced ulcerative colitis in rats. Journal of Functional Foods. 10.1016/j.jff.2019.103602
Gong Yan-Mei et al. (2026) Sinomenine ameliorates experimental autoimmune neuritis by suppressing pro-inflammatory immune components and restoring immune balance. Naunyn-Schmiedebergs Archives of Pharmacology. 10.1007/s00210-026-05151-w
Heyuan Wang et al. (2018) Protective effect of ginsenoside Rg3 on lung injury in diabetic rats. Journal of Cellular Biochemistry. 10.1002/jcb.27601
Yan Zhou et al. (2014) A diet with lactosucrose supplementation ameliorates trinitrobenzene sulfonic acid-induced colitis in rats. Food & Function. 10.1039/C4FO00381K
Yan Jiang et al. (2021) Exploring the Mechanism of Berberine Intervention in Ulcerative Colitis from the Perspective of Inflammation and Immunity Based on Systemic Pharmacology. Evidence-based Complementary and Alternative Medicine. 10.1155/2021/9970240
Jiang Hao et al. (2024) Melatonin enhances therapeutic outcomes of adipose tissue-derived mesenchymal stem cell therapy in rat osteoarthritis by reducing TNF-α and IL-1β-induced injuries. Cytotechnology. 10.1007/s10616-024-00635-0
Liu Jinyu et al. (2018) A model for anticancer surveillance was pharmacologically developed to evaluate vitality principle in breast cancer rats. Journal of Traditional Chinese Medicine. 10.1016/S0254-6272(18)30981-6
