| Field | Specification |
|---|---|
| Target | |
| Alternative names | Macrophage migration inhibitory factor ;MIF ; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: P2-A115 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Milk |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human macrophage migration inhibitory factor in cell culture supernatants, serum, plasma (heparin, EDTA) and human milk. 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.
Kit Components
| Anti-Human MIF Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human MIF Standard | 2 vials, 10 ng/tube |
| Human MIF 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 | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA) and human milk |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <20 pg/mL |
| Standard | Recombinant human Macrophage migration inhibitory factor (Pro2–Ala115), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.0–4.5% (3 samples, n = 16 each) |
| Inter-assay CV | 4.6–6.3% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:10. In the supplier’s tests, serum and plasma samples measured 5–13 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.035; 156 = 0.111; 312 = 0.196; 625 = 0.307; 1250 = 0.503; 2500 = 0.894; 5000 = 1.447; 10000 = 2.035. 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 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.067 | 0.161 | 0.272 | 0.468 | 0.859 | 1.412 | 2.0 | 2.432 |
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) | 280 | 1655 | 4559 | 305 | 1694 | 4885 |
| Standard deviation | 12.04 | 66.2 | 206.35 | 16.47 | 77.92 | 307.75 |
| CV (%) | 4.3% | 4% | 4.5% | 5.4% | 4.6% | 6.3% |
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.
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Tao Wang et al. (2026) Pancancer Fine-Mapping of Mutational Intolerance Identifies CHEK1 as an Immunosuppressive Driver in Lung Adenocarcinoma. Advanced Science. 10.1002/advs.202521265
Li Zhuang et al. (2022) Autophagy-based unconventional secretion of HMGB1 in glioblastoma promotes chemosensitivity to temozolomide through macrophage M1-like polarization. Journal of Experimental & Clinical Cancer Research. 10.1186/s13046-022-02291-8
Zhu Wei et al. (2023) Acetate attenuates hyperoxaluria-induced kidney injury by inhibiting macrophage infiltration via the miR-493-3p/MIF axis. Communications Biology. 10.1038/s42003-023-04649-w
Qiu Peng et al. (2022) Potential Clinical Value of Biomarker-Guided Emergency Triage for Thoracic Aortic Dissection. Frontiers in Cardiovascular Medicine. 10.3389/fcvm.2021.777327
Ruyi Li et al. (2020) The Role of Macrophage Migration Inhibitory Factor (MIF) in Asthmatic Airway Remodeling. Allergy Asthma & Immunology Research. 10.4168/aair.2021.13.1.88
Jue Tu et al. (2016) Chitosan nanoparticles reduce LPS-induced inflammatory reaction via inhibition of NF-κB pathway in Caco-2 cells. International Journal of Biological Macromolecules. 10.1016/j.ijbiomac.2016.02.015
Lifeng Qin et al. (2021) MIF promoter polymorphism increases peripheral blood expression levels, contributing to increased susceptibility and poor prognosis in hepatocellular carcinoma. Oncology Letters. 10.3892/ol.2021.12810
Li-Rong Lin et al. (2016) Macrophage migration inhibitory factor as a novel cerebrospinal fluid marker for neurosyphilis among HIV-negative patients. Clinica Chimica ACTA. 10.1016/j.cca.2016.10.018
Zhentong Liu et al. (2023) Retrospective study on the correlation between serum MIF level and the condition and prognosis of patients with traumatic head injury. PeerJ. 10.7717/peerj.15933
Mehmet Calan et al. (2016) A possible link between luteinizing hormone and macrophage migration inhibitory factor levels in polycystic ovary syndrome. Endocrine Research. 10.3109/07435800.2015.1135442
