| Field | Specification |
|---|---|
| Target | |
| Alternative names | Interleukin-6; Il6 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: F25-T211 |
| 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 one-step sandwich ELISA quantifies mouse interleukin-6 (gene Il6) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). 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.
Interleukin-6 (IL-6) is a multifunctional cytokine released by macrophages, dendritic cells and many other cells during infection and tissue injury. It binds IL-6R and then the signaling subunit gp130, activating JAK–STAT3 through classic or trans-signaling. IL-6 drives the acute phase response, B-cell differentiation and Th17 development, and it is one of the most widely measured inflammatory markers.
Kit Components
| Anti-Mouse Il6 Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Mouse Il6 Standard | 2 × 10 ng/tube |
| HRP Linked Anti-Mouse Il6 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): microplate reader for 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.
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 | <1 pg/mL |
| Standard | Recombinant mouse Il6 (Phe25–Thr211), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.4–6.1% (3 samples, n = 16 each) |
| Inter-assay CV | 6.5–7.8% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.022; 15.6 = 0.055; 31.2 = 0.110; 62.5 = 0.183; 125 = 0.317; 250 = 0.588; 500 = 1.068; 1000 = 1.829. Run a new standard curve on every plate.
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 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.022 | 0.055 | 0.110 | 0.183 | 0.317 | 0.588 | 1.068 | 1.829 |
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) | 40 | 168 | 445 | 41 | 172 | 459 |
| Standard deviation | 2.16 | 10.24 | 24.47 | 3.15 | 13.41 | 29.83 |
| CV (%) | 5.4% | 6.1% | 5.5% | 7.7% | 7.8% | 6.5% |
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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