| Field | Specification |
|---|---|
| Target | |
| Alternative names | Monocyte differentiation antigen CD14; Myeloid cell-specific leucine-rich glycoprotein; CD14 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: CHO; Immunogen sequence: T20-C352 |
| 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 human monocyte differentiation antigen CD14 in cell culture supernatants, serum and plasma (heparin, EDTA, citrate ). 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.
CD14 is a pattern-recognition co-receptor on monocytes and macrophages that, together with LBP, captures bacterial lipopolysaccharide (LPS) and delivers it to the TLR4/MD-2 complex. This triggers NF-κB activation and inflammatory cytokine release, and CD14 is also a standard marker for monocytes in flow cytometry.
Kit Components
| Anti-Human CD14 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human CD14 Standard | 2 vials, 10 ng/tube |
| Human CD14 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 and plasma (heparin, EDTA, citrate ) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <50 pg/mL |
| Standard | Recombinant human CD14 (Thr20–Cys352), expressed in CHO |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.5–5.1% (3 samples, n = 16 each) |
| Inter-assay CV | 5.8–7.4% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:4000. In the supplier’s tests, serum and plasma samples measured about 4800 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.033; 156 = 0.095; 312 = 0.155; 625 = 0.259; 1250 = 0.42; 2500 = 0.759; 5000 = 1.264; 10000 = 2.024. 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. | 0.033 | 0.095 | 0.155 | 0.259 | 0.42 | 0.759 | 1.264 | 2.024 |
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) | 310 | 1057 | 4116 | 312 | 1164 | 4732 |
| Standard deviation | 13.95 | 49.7 | 209.92 | 18.1 | 75.66 | 350.2 |
| CV (%) | 4.5% | 4.7% | 5.1% | 5.8% | 6.5% | 7.4% |
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.
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Bhaskaran N. et al. (2021) Oral immune dysfunction is associated with the expansion of FOXP3+PD-1+Amphiregulin+ T cells during HIV infection. Nature Communications. 10.1038/s41467-021-25340-w
S Jayaraman et al. (2026) Enrichment of Candida associated with dysbiosis contributes to mucosal CD4+FOXP3+ regulatory T cell accrual and their dysfunction in aging. Frontiers in Immunology. 10.3389/fimmu.2026.1714595
