| Field | Specification |
|---|---|
| Target | |
| Alternative names | Macrophage metalloelastase;MME;3.4.24.65;Macrophage elastase;ME;hME;Matrix metalloproteinase-12;MMP-12;MMP12;HME; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: L17-C470 |
| 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 macrophage metalloelastase (gene MMP12) 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.
Macrophage metalloelastase (MMP-12) is a matrix metalloproteinase produced mainly by macrophages that breaks down elastin and other matrix proteins. It is widely studied in lung tissue injury and remodeling, especially in models of emphysema and chronic inflammation.
Kit Components
| Anti-Human MMP12 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human MMP12 Standard | 2 vials, 10 ng/tube |
| Human MMP12 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 | 62.5 pg/mL–4,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human MMP12 (Leu17–Cys470), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.1–6.4% (3 samples, n = 16 each) |
| Inter-assay CV | 5.8–6.8% (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 about 500 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.049; 62.5 = 0.134; 125 = 0.205; 250 = 0.309; 500 = 0.51; 1000 = 0.838; 2000 = 1.393; 4000 = 2.082. 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 | 62.5 | 125 | 250 | 500 | 1000 | 2000 | 4000 |
| O.D. | 0.049 | 0.158 | 0.26 | 0.461 | 0.789 | 1.244 | 1.743 | 2.163 |
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) | 201 | 499 | 1029 | 197 | 507 | 1084 |
| Standard deviation | 12.86 | 20.45 | 59.68 | 13.39 | 30.42 | 62.87 |
| CV (%) | 6.4% | 4.1% | 5.8% | 6.8% | 6% | 5.8% |
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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Amor Melina et al. (2023) Genetic deletion of MMP12 ameliorates cardiometabolic disease by improving insulin sensitivity, systemic inflammation, and atherosclerotic features in mice. Cardiovascular Diabetology. 10.1186/s12933-023-02064-3
Eleni Papakonstantinou et al. (2016) Acute Exacerbations of COPD Are Associated With Increased Expression of Heparan Sulfate and Chondroitin Sulfate in BAL. Chest. 10.1378/chest.14-2868
Jiansheng Li et al. (2021) A chinese herbal formula ameliorates COPD by inhibiting the inflammatory response via downregulation of p65, JNK, and p38. Phytomedicine. 10.1016/j.phymed.2021.153475
Sarra Bchir et al. (2016) Concomitant elevations of MMP-9, NGAL, proMMP-9/NGAL and neutrophil elastase in serum of smokers with chronic obstructive pulmonary disease. Journal of Cellular and Molecular Medicine. 10.1111/jcmm.13057
Bchir Sarra et al. (2021) Impact of cathepsin D activity and C224T polymorphism (rs17571) on chronic obstructive pulmonary disease: correlations with oxidative and inflammatory markers. Clinical and Experimental Medicine. 10.1007/s10238-021-00692-1
Lin Du et al. (2022) Identification of CCL20 and LCN2 as Efficient Serological Tools for Detection of Hepatocellular Carcinoma. Disease Markers. 10.1155/2022/7758735
Wendong Hao et al. (2019) Expressions of MMP-12, TIMP-4, and Neutrophil Elastase in PBMCs and Exhaled Breath Condensate in Patients with COPD and Their Relationships with Disease Severity and Acute Exacerbations. Journal of Immunology Research. 10.1155/2019/7142438
T. Narimiya et al. (2017) Orthodontic tensile strain induces angiogenesis via type IV collagen degradation by matrix metalloproteinase-12. Journal of Periodontal Research. 10.1111/jre.12453
Sarra Bchir et al. (2017) MMP-3 (–1171 5A/6A; Lys45Glu) variants affect serum levels of matrix metalloproteinase (MMP)-3 and correlate with severity of COPD: A study of MMP-3, MMP-7 and MMP-12 in a Tunisian population. Journal of Gene Medicine. 10.1002/jgm.2999
Kazuhiko Kobayashi et al. (2017) Elastin‑derived peptides are involved in the processes of human temporomandibular disorder by inducing inflammatory responses in synovial cells. Molecular Medicine Reports. 10.3892/mmr.2017.7012
