| Field | Specification |
|---|---|
| Target | |
| Alternative names | Metalloproteinase inhibitor 1;Tissue inhibitor of metalloproteinases 1;TIMP-1;Timp1;Timp-1; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: C24-A217 |
| 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 rat metalloproteinase inhibitor 1 (gene Timp1) 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.
TIMP-1 is a secreted tissue inhibitor of metalloproteinases that binds the catalytic zinc site of many matrix metalloproteinases, including MMP-9, and blocks their activity. It also has growth factor–like, MMP-independent effects on cell survival and proliferation. TIMP-1 is studied in extracellular matrix remodeling, fibrosis, inflammation and cancer.
Kit Components
| Anti-Rat Timp1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Rat Timp1 Standard | 2 vials, 10 ng/tube |
| Rat Timp1 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 | Rat |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Sensitivity | <3 pg/mL |
| Standard | Recombinant rat Timp1 (Cys24–Ala217), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.2–7.2% (3 samples, n = 16 each) |
| Inter-assay CV | 5.2–8.2% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:50. In the supplier’s tests, serum and plasma samples measured 5920–49680 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.031; 31.2 = 0.125; 62.5 = 0.195; 125 = 0.316; 250 = 0.566; 500 = 1.008; 1000 = 1.651; 2000 = 2.781. 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 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.031 | 0.125 | 0.195 | 0.316 | 0.566 | 1.008 | 1.651 | 2.781 |
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) | 59 | 252 | 889 | 61 | 252 | 895 |
| Standard deviation | 4.24 | 13.1 | 45.33 | 5 | 13.35 | 46.54 |
| CV (%) | 7.2% | 5.2% | 5.2%% | 8.2% | 5.3% | 5.2% |
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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Lan Liu et al. (2021) Effective-components combination improves airway remodeling in COPD rats by suppressing M2 macrophage polarization via the inhibition of mTORC2 activity. Phytomedicine. 10.1016/j.phymed.2021.153759
Jing Wang et al. (2020) Exposure to Air Pollution Exacerbates Inflammation in Rats with Preexisting COPD. Mediators of Inflammation. 10.1155/2020/4260204
Jiansheng Li et al. (2020) Effective-component compatibility of Bufei Yishen formula II inhibits mucus hypersecretion of chronic obstructive pulmonary disease rats by regulating EGFR/PI3K/mTOR signaling. Journal of Ethnopharmacology. 10.1016/j.jep.2020.112796
Ural Cemre et al. (2023) The renoprotective effects of taurine against diabetic nephropathy via the p38 MAPK and TGF-β/Smad2/3 signaling pathways. Amino Acids. 10.1007/s00726-023-03342-w
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Yu He et al. (2018) Mahuang decoction mitigates airway inflammation and regulates IL-21/STAT3 signaling pathway in rat asthma model. Journal of Ethnopharmacology. 10.1016/j.jep.2018.06.011
Le Shi et al. (2016) Traditional Chinese Medicine Formula “Xiaofeng granules” suppressed gouty arthritis animal models and inhibited the proteoglycan degradation on chondrocytes induced by monosodium urate. Journal of Ethnopharmacology. 10.1016/j.jep.2016.06.008
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