| Field | Specification |
|---|---|
| Target | |
| Alternative names | Collagenase 3;3.4.24.-;Matrix metalloproteinase-13;MMP-13;MMP13; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: L20-C471 |
| 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 collagenase 3 (gene MMP13) in cell culture supernatants, serum and plasma ( heparin). 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.
Collagenase 3 (MMP-13) is a matrix metalloproteinase that degrades fibrillar collagens, with the highest activity toward type II collagen of cartilage, and it also cleaves aggrecan and other matrix proteins. It is needed for normal bone development and is widely studied in cartilage breakdown in osteoarthritis.
Kit Components
| Anti-Human MMP13 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human MMP13 Standard | 2 vials, 10 ng/tube |
| Human MMP13 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) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <5 pg/mL |
| Standard | Recombinant human MMP13 (Leu20–Cys471), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 6.4–7.0% (3 samples, n = 16 each) |
| Inter-assay CV | 7.5–8.1% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.025; 156 = 0.085; 312 = 0.164; 625 = 0.345; 1250 = 0.602; 2500 = 1.078; 5000 = 1.536; 10000 = 2.011. 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. | 0.025 | 0.085 | 0.164 | 0.345 | 0.602 | 1.078 | 1.536 | 2.011 |
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) | 219 | 1742 | 4482 | 203 | 1816 | 4413 |
| Standard deviation | 14.01 | 116.71 | 313.74 | 16.4%2 | 146.4%8 | 357.45 |
| CV (%) | 6.4% | 6.7% | 7% | 7.5% | 8% | 8.1% |
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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Nazir M. Khan et al. (2017) Wogonin, a plant derived small molecule, exerts potent anti-inflammatory and chondroprotective effects through the activation of ROS/ERK/Nrf2 signaling pathways in human Osteoarthritis chondrocytes. Free Radical Biology and Medicine. 10.1016/j.freeradbiomed.2017.02.041
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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
Dong Yang et al. (2022) Epigallocatechin-3-O-gallate promotes extracellular matrix and inhibits inflammation in IL-1β stimulated chondrocytes by the PTEN/miRNA-29b pathway. Pharmaceutical Biology. 10.1080/13880209.2022.2039722
Riwon Hong et al. (2017) Anti-inflammatory and anti-arthritic effects of the ethanolic extract of Aralia continentalis Kitag. in IL-1β-stimulated human fibroblast-like synoviocytes and rodent models of polyarthritis and nociception. Phytomedicine. 10.1016/j.phymed.2017.10.016
Weidong Zhu et al. (2022) Glaucocalyxin A Attenuates IL-1β-Induced Inflammatory Response and Cartilage Degradation in Osteoarthritis Chondrocytes via Inhibiting the Activation of NF-κB Signaling Pathway. Disease Markers. 10.1155/2022/6516246
