| Field | Specification |
|---|---|
| Target | |
| Alternative names | CD27 antigen; CD27L receptor; T-cell activation antigen CD27; T14; Tumor necrosis factor receptor superfamily member 7; CD antigen CD27; CD27; TNFRSF7 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: T21-I192 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This single-incubation sandwich ELISA quantifies human CD27 antigen in cell culture supernatants and serum. The strips are coated with an anti-tag antibody. Samples and standards are added to the wells together with the kit’s antibody cocktail (Antibody A and Antibody B); the resulting antibody–target complex is held on the plate through the tag. After washing, TMB substrate is added and the HRP label on the detection antibody converts it to a colored product in proportion to the bound target. The reaction is stopped and the plate is read at 450 nm. The supplier markets the Quick format as a 90-minute assay.
CD27 is a TNF receptor superfamily co-stimulatory receptor on T cells, NK cells and B cells that binds CD70. CD70–CD27 signaling supports T-cell activation, survival and memory formation, and CD27 is a commonly used marker for memory B cells and T-cell differentiation states.
Kit Components
| Anti-tag Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human CD27 Standard | 2 × 10 ng/tube |
| Human CD27 Antibody A | 3 mL |
| Human CD27 Antibody B | 3 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 | Human |
|---|---|
| Sample types | Cell culture supernatants and serum |
| Assay range | 62.5 pg/mL–4,000 pg/mL |
| Sensitivity | <1 pg/mL |
| Standard | Recombinant human CD27 (Thr21–Ile192), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 3.6–6.5% (3 samples, n = 16 each) |
| Inter-assay CV | 5.7–6.1% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.064; 62.5 = 0.174; 125 = 0.272; 250 = 0.452; 500 = 0.782; 1,000 = 1.367; 2,000 = 2.134; 4,000 = 2.496. 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 | 62.5 | 125 | 250 | 500 | 1,000 | 2,000 | 4,000 |
| O.D. | 0.064 | 0.174 | 0.272 | 0.452 | 0.782 | 1.367 | 2.134 | 2.496 |
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) | 105 | 210 | 1426 | 107 | 541 | 1466 |
| Standard deviation | 6.83 | 22.95 | 51.34 | 6.21 | 30.84 | 89.43 |
| CV (%) | 6.5% | 4.5% | 3.6% | 5.8% | 5.7% | 6.1% |
Kit components
Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml 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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