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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
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL)
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Sensitivity
4.5 pg/mL
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Assay Range
37.5 - 2,400 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
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Specificity
Natural and recombinant rat TIMP-1
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Rat TIMP-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure rat TIMP-1 levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant rat TIMP-1 and antibodies raised against the recombinant protein. Natural rat TIMP-1 showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural rat 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 | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 122 | 443 | 1307 | 132 | 445 | 1277 |
| Standard Deviation | 4.5 | 14 | 34 | 9.6 | 27 | 85 |
| CV% | 3.7 | 3.2 | 2.6 | 7.3 | 6.1 | 6.7 |
Recovery
The recovery of rat TIMP-1 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=5) | 106 | 93-119 |
| EDTA Plasma (n=7) | 99 | 82-115 |
| Serum (n=7) | 98 | 84-120 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with rat TIMP-1 were diluted with Calibrator Diluent and then assayed.
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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