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.
Human TIMP-4 Quantikine ELISA Kit
R&D Systems | Catalog # DTM400
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Human Milk (10 µL)
Sensitivity
10 pg/mL
Assay Range
78.1-5000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Human Milk)
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Product Summary for Human TIMP-4 Quantikine ELISA Kit
The Quantikine Human TIMP-4 Immunoassay is a 4.5 hour solid phase ELISA designed to measure TIMP-4 levels in cell culture supernates, serum, plasma, and human milk. It contains Sf 21-expressed recombinant human TIMP-4 and antibodies raised against the recombinant protein. This kit detects recombinant human TIMP-4 in both free and MMP-bound forms. Natural human TIMP-4 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human TIMP-4.
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-4. This kit detects recombinant human TIMP-4 in both free and MMP-bound forms.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.
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
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 252 | 793 | 1476 | 232 | 708 | 1377 |
| Standard Deviation | 11.0 | 46.9 | 77.8 | 20.8 | 45.8 | 96.4 |
| CV% | 4.4 | 5.9 | 5.3 | 9.0 | 6.5 | 7.0 |
EDTA Plasma, Heparin Plasma, Human Milk, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 235 | 768 | 1516 | 220 | 712 | 1414 |
| Standard Deviation | 13.2 | 21.5 | 70.4 | 20.2 | 48.2 | 87.6 |
| CV% | 5.6 | 2.8 | 4.6 | 9.2 | 6.8 | 6.2 |
Recovery for Human TIMP-4 Quantikine ELISA Kit
The recovery of TIMP-4 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Supernates (n=4) | 96 | 87-105 |
| EDTA Plasma (n=4) | 95 | 87-113 |
| Heparin Plasma (n=4) | 93 | 86-104 |
| Serum (n=4) | 96 | 86-103 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of TIMP-4 were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human TIMP-4 Quantikine ELISA Kit
Human TIMP-4 ELISA Calibrator Diluent RD6-31 Standard Curve
Human TIMP-4 ELISA Calibrator Diluent RD6-31 (1X) 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-4
Long Name
Tissue Inhibitors of Metalloproteinases 4
Alternate Names
TIMP4
Gene Symbol
TIMP4
Additional TIMP-4 Products
Product Documents for Human TIMP-4 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-4 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-4 Quantikine ELISA Kit
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Protocols
View specific protocols for Human TIMP-4 Quantikine ELISA Kit (DTM400):
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.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. 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
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