| Field | Specification |
|---|---|
| Target | |
| Alternative names | Metalloproteinase inhibitor 2;CSC-21K;Tissue inhibitor of metalloproteinases 2;TIMP-2;TIMP2; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: CHO; Immunogen sequence: C27-P220 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Saliva |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human metalloproteinase inhibitor 2 (gene TIMP2) in cell culture supernatants, serum, plasma (heparin, EDTA) and saliva. 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.
TIMP-2 is a secreted tissue inhibitor of metalloproteinases that inhibits a broad range of MMPs. It also has a special role in activating pro-MMP-2 at the cell surface together with MT1-MMP (MMP-14). TIMP-2 is studied in extracellular matrix turnover, angiogenesis, kidney injury and tumor biology.
Kit Components
| Anti-Human TIMP2 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human TIMP2 Standard | 2 vials, 10 ng/tube |
| Human TIMP2 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, plasma (heparin, EDTA) and saliva |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <2 pg/mL |
| Standard | Recombinant human TIMP2 (Cys27–Pro220), expressed in CHO |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.3–7.4% (3 samples, n = 16 each) |
| Inter-assay CV | 6.8–8.3% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:100. In the supplier’s tests, serum and plasma samples measured 260 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.03; 156 = 0.08; 312 = 0.128; 625 = 0.231; 1250 = 0.405; 2500 = 0.77; 5000 = 1.336; 10000 = 2.165. 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. | 0.03 | 0.08 | 0.128 | 0.231 | 0.405 | 0.77 | 1.336 | 2.165 |
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) | 249 | 1786 | 4399 | 239 | 1654 | 4356 |
| Standard deviation | 17.43 | 94.65 | 325.52 | 16.73 | 112.47 | 361.54 |
| CV (%) | 6.4% | 5.3% | 7.4% | 7% | 6.8% | 8.3% |
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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Vincenzo Di Stefano et al. (2022) Metalloproteinases and Tissue Inhibitors in Generalized Myasthenia Gravis. A Preliminary Study. Brain Sciences. 10.3390/brainsci12111439
Cosentino Giuseppe et al. (2021) Expression pattern of matrix metalloproteinases-2 and -9 and their tissue inhibitors in patients with chronic inflammatory demyelinating polyneuropathy. Neurological Sciences. 10.1007/s10072-021-05314-y
E. Hopps et al. (2014) Study of the Correlations among Some Parameters of the Oxidative Status, Gelatinases, and Their Inhibitors in a Group of Subjects with Metabolic Syndrome. Mediators of Inflammation. 10.1155/2014/510619
Wen-jing Zhang et al. (2019) MMP-2, MMP-9, TIMP-1, and TIMP-2 in the Peripheral Blood of Patients with Differentiated Thyroid Carcinoma. Cancer Management and Research. 10.2147/CMAR.S233776
Caimi G. et al. (2015) Behaviour of the plasma concentration of gelatinases and their tissue inhibitors in subjects with venous leg ulcers. Clinical Hemorheology and Microcirculation. 10.3233/CH-141863
Hopps Eugenia et al. (2016) Gelatinases and their tissue inhibitors in a group of subjects with obstructive sleep apnea syndrome. Clinical Hemorheology and Microcirculation. 10.3233/CH-151928
Eugenia Hopps et al. (2013) Gelatinases and Their Tissue Inhibitors in a Group of Subjects With Metabolic Syndrome. Journal of Investigative Medicine. 10.2310/JIM.0b013e318294e9da
Kim Kyoung Soo et al. (2011) Implication of MMP-9 and urokinase plasminogen activator (uPA) in the activation of pro-matrix metalloproteinase (MMP)-13. Rheumatology International. 10.1007/s00296-011-2095-4
Yu Xiaoping et al. (2010) Anti-angiogenic genistein inhibits VEGF-induced endothelial cell activation by decreasing PTK activity and MAPK activation. Medical Oncology. 10.1007/s12032-010-9770-2
