| Field | Specification |
|---|---|
| Target | |
| Alternative names | Leukemia inhibitory factor;LIF;Differentiation-stimulating factor;D factor;Lif; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S24-F203 |
| 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 leukemia inhibitory factor (gene Lif) in cell culture supernatants, serum and plasma (heparin, EDTA). 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.
Leukemia inhibitory factor (LIF) is an IL-6 family cytokine that signals through a receptor made of LIFR and gp130, activating JAK–STAT3 signaling. It maintains self-renewal of mouse embryonic stem cells in culture and is essential for embryo implantation. LIF also acts on neurons, bone, muscle and immune cells and is widely used in stem cell culture and cytokine research.
Kit Components
| Anti-Mouse Lif Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Lif Standard | 2 vials, 10 ng/tube |
| Mouse Lif 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) |
| Assay range | 7.8 pg/mL–500 pg/mL |
| Sensitivity | <2 pg/mL |
| Standard | Recombinant mouse Lif (Ser24–Phe203), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.3–7.2% (3 samples, n = 16 each) |
| Inter-assay CV | 4.4–8.9% (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 below the lowest standard.
Example standard curve (OD450, pg/mL): 0 = 0.029; 7.8 = 0.071; 15.6 = 0.131; 31.3 = 0.265; 62.5 = 0.507; 125 = 0.837; 250 = 1.315; 500 = 1.81. 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. | 0.029 | 0.071 | 0.131 | 0.265 | 0.507 | 0.837 | 1.315 | 1.81 |
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) | 16 | 70 | 272 | 15 | 76 | 278 |
| Standard deviation | 0.7 | 3.01 | 19.58 | 0.72 | 3.34 | 24.74 |
| CV (%) | 4.4% | 4.3% | 7.2% | 4.8% | 4.4% | 8.9% |
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.
Activating reagent preparation
LIF is mostly contained as inactive form in samples, please activate it before assay. Don't activate recombinant LIF. Solution A: 1N HCI: add 8.33ml of 12N HCI into 91.67ml of H2O. Solution B: 1.2N NaOH/0.5M HEPES: add 12ml of 10N NaOH and 11.9g HEPES into 75ml of H2O, add H2O to adjust volume to 100ml.
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Jiawei Zhang et al. (2019) Low-level laser therapy 810-nm up-regulates macrophage secretion of neurotrophic factors via PKA-CREB and promotes neuronal axon regeneration in vitro. Journal of Cellular and Molecular Medicine. 10.1111/jcmm.14756
