| Field | Specification |
|---|---|
| Target | |
| Alternative names | Leukemia inhibitory factor; LIF; Differentiation-stimulating factor; D factor; Melanoma-derived LPL inhibitor; MLPLI; Emfilermin; HILDA |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S23-F202 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Urine |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human leukemia inhibitory factor (gene LIF) in cell culture supernatants, serum, plasma (heparin, EDTA) and urine. 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-Human LIF Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human LIF Standard | 2 × 10 ng/tube |
| Human 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 |
| 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.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA) and urine |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Sensitivity | <8 pg/mL |
| Standard | Recombinant human LIF (Ser23–Phe202), expressed in E. coli |
| Cross-reactivity | No significant cross-reactivity or interference with LIF analogs in supplier testing; untested analogs cannot be excluded. |
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. | 0.033 | 0.111 | 0.179 | 0.301 | 0.574 | 0.89 | 1.5 | 2.085 |
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) | 23 | 40 | 341 | 30 | 54 | 431 |
| Standard deviation | 0.6 | 2.14 | 18.96 | 0.79 | 1.73 | 18.02 |
| CV(%) | 2.6% | 5.3% | 5.6% | 2.6% | 3.2% | 4.2% |
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.
Sample activation procedure
Activate the sample
Serum, plasma (EDTA): add activating reagent pro rata, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH 7.0-7.6.
It is unnecessary to activate the Cell culture supernate.
It is unnecessary to activate the recombinant LIF.
Sample was diluted partly after adding activating reagent, so please pay attention to this when calculate target protein concentration.
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Kabir Ahluwalia et al. (2023) Polarized RPE Secretome Preserves Photoreceptors in Retinal Dystrophic RCS Rats. Cells. 10.3390/cells12131689
