| Field | Specification |
|---|---|
| Target | |
| Alternative names | A disintegrin and metalloproteinase with thrombospondin motifs 4;ADAM-TS 4;ADAM-TS4;ADAMTS-4;3.4.24.82;ADMP-1;Aggrecanase-1;ADAMTS4;KIAA0688;UNQ769/PRO1563; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: CHO; Immunogen sequence: F213-C685 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human A disintegrin and metalloproteinase with thrombospondin motifs 4 (gene ADAMTS4) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). 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.
ADAMTS4 (aggrecanase-1) is a secreted metalloprotease that cleaves aggrecan and other cartilage proteoglycans. It contributes to cartilage turnover and is studied as a mediator of cartilage breakdown in osteoarthritis.
Kit Components
| Anti-Human ADAMTS4 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human ADAMTS4 Standard | 2 vials, 40 ng/tube |
| Human ADAMTS4 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, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 625 pg/mL–40,000 pg/mL |
| Sensitivity | <20 pg/mL |
| Standard | Recombinant human ADAMTS4 (Phe213–Cys685), expressed in CHO |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.1–5.6% (3 samples, n = 16 each) |
| Inter-assay CV | 4.3–7.1% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum and plasma samples measured below the lowest standard.
Example standard curve (OD450, ng/mL): 0 = 0.064; 0.625 = 0.16; 1.25 = 0.222; 2.5 = 0.308; 5 = 0.522; 10 = 0.921; 20 = 1.448; 40 = 2.465. 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 (ng/ml) | 0 | 0.625 | 1.25 | 2.5 | 5 | 10 | 20 | 40 |
| O.D. | 0.064 | 0.16 | 0.222 | 0.308 | 0.522 | 0.921 | 1.448 | 2.465 |
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) | 1127 | 4290 | 29063 | 1210 | 4672 | 29313 |
| Standard deviation | 46.2 | 184.47 | 1627.52 | 71.39 | 200.89 | 2081.22 |
| CV (%) | 4.1% | 4.3% | 5.6% | 5.9% | 4.3% | 7.1% |
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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Weinmann Daniela et al. (2016) Galectin-3 Induces a Pro-degradative/inflammatory Gene Signature in Human Chondrocytes, Teaming Up with Galectin-1 in Osteoarthritis Pathogenesis. Scientific Reports. 10.1038/srep39112
Toegel et al. (2016) Galectin-1 Couples Glycobiology to Inflammation in Osteoarthritis through the Activation of an NF-κB–Regulated Gene Network. Journal of Immunology. 10.4049/jimmunol.1501165
Xinxin Liu et al. (2020) Sodium butyrate protects against oxidative stress in human nucleus pulposus cells via elevating PPARγ-regulated Klotho expression. International Immunopharmacology. 10.1016/j.intimp.2020.106657
