| Field | Specification |
|---|---|
| Target | |
| Alternative names | Tumor necrosis factor ligand superfamily member 6;Apoptosis antigen ligand;APTL;CD95 ligand;CD95-L;Fas antigen ligand;Fas ligand;FasL;CD178;Tumor necrosis factor ligand superfamily member 6, membrane form;Tumor necrosis factor ligand superfamily member 6, soluble form;Receptor-binding FasL ectodomain;Soluble Fas ligand;sFasL;ADAM10-processed FasL form;APL;FasL intracellular domain;FasL ICD;SPPL2A-processed FasL form;SPA;FASLG;APT1LG1, CD95L, FASL, TNFSF6; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: CHO; Immunogen sequence: P134-L281 |
| 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 single-incubation sandwich ELISA quantifies human tumor necrosis factor ligand superfamily member 6 (gene FASLG) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). 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.
Fas ligand (FasL, CD95L) is a TNF family cytokine that triggers apoptosis by binding its receptor Fas. It is expressed by cytotoxic T cells and NK cells and helps them kill target cells; it also drives activation-induced death of T cells, which limits immune responses. FasL is studied in immune regulation, immune privilege and cancer immunology.
Kit Components
| Anti-tag Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human FASLG Standard | 2 × 10 ng/tube |
| Human FASLG Antibody A | 3 mL |
| Human FASLG 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, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Sensitivity | <2 pg/mL |
| Standard | Recombinant human FASLG (Pro134–Leu281), expressed in CHO |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 3.3–7.2% (3 samples, n = 16 each) |
| Inter-assay CV | 2.8–6.4% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.014; 15.6 = 0.053; 31.2 = 0.085; 62.5 = 0.152; 125 = 0.279; 250 = 0.551; 500 = 1.033; 1000 = 2.059. 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 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.008 | 0.114 | 0.196 | 0.379 | 0.740 | 1.371 | 2.121 | 2.693 |
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) | 36 | 186 | 437 | 34 | 189 | 419 |
| Standard deviation | 2.19 | 9.67 | 28.84 | 2.1 | 12.47 | 33.1 |
| CV (%) | 6.1% | 5.2% | 6.6% | 6.2% | 6.6% | 7.9% |
Kit components
Description|Quantity 96-well plate|1 lyophilized Standard| Biotinylated detection antibody (50x)|100 ul Avidin-Biotin-Peroxidase Complex (30x)|400 µl Sample diluent|30 ml Antibody diluent|12 ml Avidin-Biotin-Peroxidase Diluent|12 ml Wash Buffer (25x)|20 ml Color Developing Reagent (TMB)|10 ml Stop Solution|10 ml Plate Sealers|4Materials 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.
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