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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
3.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL)
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Sensitivity
0.08 ng/mL
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Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva)
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Specificity
Natural and recombinant human TIMP-1
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
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Interference
Interference observed with 1 or more available related molecules.
Control Available
QC173, Quantikine Immunoassay Control Set 651 for Human TIMP-1 -
Product Summary
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.
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, Serum, EDTA Plasma, Heparin Plasma, Saliva | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 0.48 | 1.27 | 6.95 | 0.51 | 1.28 | 6.9 |
| Standard Deviation | 0.02 | 0.05 | 0.35 | 0.02 | 0.05 | 0.34 |
| CV% | 4.2 | 3.9 | 5 | 3.9 | 3.9 | 4.9 |
Recovery
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.
Preparation and Storage
Background: TIMP-1
TIMPs-1 through -4 regulate the activity of zinc metalloproteases known as MMPs, ADAMs and ADAMTSs. Structurally, TIMPs contain two domains. The N-terminal domain binds to the active site of mature metalloproteases via a 1:1 non-covalent interaction, blocking access of substrates to the catalytic site. In addition, The C-terminal domain of TIMP-1 and TIMP-2 binds to the hemopexin- like domain of pro-MMP-9 and pro-MMP-2, respectively. The latter binding is essential for the cell surface activation of MMP-2 by MMP-14.
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