TIMP-1 (tissue inhibitor of metalloproteinase-1) is a secreted glycoprotein that blocks metalloproteinase (MMP) activity. It circulates in complex with various MMPs to maintain them in an inactive state. It is produced by fibroblasts, osteoblasts, endothelial cells, granulosa cells, dendritic cells, vascular smooth muscle cells, adipocytes, and monocytes. TIMP-1 also protects against apoptosis in B cells, stimulates erythropoiesis and BFU-E expansion, both stimulates and inhibits tumor growth, inhibits angiogenesis, and stimulates gonadal steroidogenesis in conjuction with Cathepsin L.
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Human TIMP-1 Quantikine ELISA Kit
R&D Systems | Catalog # DTM100
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Key Product Details
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (10 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Saliva (10 µL)
Sensitivity
0.08 ng/mL
Assay Range
0.156-10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva)
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Product Summary for Human TIMP-1 Quantikine ELISA Kit
The Quantikine Human TIMP-1 Immunoassay is a 3.5 hour solid phase ELISA designed to measure TIMP-1 concentrations in cell culture supernates, serum, plasma, and saliva. It contains NS0-expressed recombinant human TIMP-1 and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human TIMP-1 accurately. Results obtained using natural TIMP-1 showed linear curves that were parallel to the standard curve obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural TIMP-1.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human TIMP-1
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not tested.
Interference
Interference observed with 1 or more available related molecules.
Sample Values
Serum/Plasma/Saliva - Samples from apparently healthy volunteers were evaluated for the
presence of human TIMP-1 in this assay. No medical histories were available for the donors
used in this study.
Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and assayed for levels of natural human TIMP-1.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Serum (n=60) | 190 | 87-524 | 72.1 |
| Heparin plasma (n=60) | 84 | 39-279 | 33.1 |
| EDTA plasma (n=60) | 98 | 44-304 | 35.4 |
| Saliva (n=4) | 121 | 46-208 | 82.3 |
Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and assayed for levels of natural human TIMP-1.
| Condition | Day 1 (ng/mL) | Day 5 (ng/mL) |
| Unstimulated | 76 | 163 |
| Stimulated | 201 | 274 |
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Saliva, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (ng/mL) | 0.48 | 1.27 | 6.95 | 0.51 | 1.28 | 6.90 |
| Standard Deviation | 0.02 | 0.05 | 0.35 | 0.02 | 0.05 | 0.34 |
| CV% | 4.2 | 3.9 | 5.0 | 3.9 | 3.9 | 4.9 |
Recovery for Human TIMP-1 Quantikine ELISA Kit
The recovery of TIMP-1 spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 102 | 94-113 |
| EDTA Plasma (n=5) | 99 | 93-108 |
| Heparin Plasma (n=5) | 99 | 89-108 |
| Saliva (n=4) | 105 | 89-121 |
| Serum (n=5) | 98 | 87-102 |
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of TIMP-1 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human TIMP-1 Quantikine ELISA Kit
Human TIMP-1 ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TIMP-1
Long Name
Tissue Inhibitors of Metalloproteinases 1
Alternate Names
TIMP1
Gene Symbol
TIMP1
Additional TIMP-1 Products
Product Documents for Human TIMP-1 Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human TIMP-1 Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human TIMP-1 Quantikine ELISA Kit
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Sample Tested: SerumVerified Customer | Posted 11/25/2022
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Sample Tested: UrineVerified Customer | Posted 03/15/2022Urine was diluted 2-fold, 4-fold, and 8-fold. Two of 8 samples had a reference value below range, 5/8 samples were in range at the 4-fold dilution, and 4/8 samples were in range at the 8-fold dilution. The samples that were out of range were all BLQ. Despite some samples being BLQ at the lower dilutions, most of the samples showed good dilution linearity with %Recoveries within 80-120%.
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Sample Tested: SerumVerified Customer | Posted 06/07/2021sample dilution 1:3
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Sample Tested: Serum and PlasmaVerified Customer | Posted 07/17/2020
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Protocols
View specific protocols for Human TIMP-1 Quantikine ELISA Kit (DTM100):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process twice for a total of 3 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
- Aspirate and wash 3 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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