Rat TIMP-1 Quantikine ELISA Kit

  (4 citations)     
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Assay Procedure
Citations (4)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL)
  • Sensitivity
    4.5 pg/mL
  • Assay Range
    37.5 - 2,400 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant rat TIMP-1
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • 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.
 TIMP-1 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
  • Long Name:
    Tissue Inhibitors of Metalloproteinases 1
  • Entrez Gene IDs:
    7076 (Human); 21857 (Mouse); 116510 (Rat)
  • Alternate Names:
    CLGI; Collagenase inhibitor; collagenase inhibitor); EPATIMP-1; EPO; erythroid potentiating activity; Erythroid-potentiating activity; Fibroblast collagenase inhibitor; FLJ90373; HCI; metalloproteinase inhibitor 1; TIMP metallopeptidase inhibitor 1; TIMP1; TIMP-1; TIMPtissue inhibitor of metalloproteinase 1 (erythroid potentiating activity; Tissue inhibitor of metalloproteinases 1
Related Research Areas
Assay Procedure
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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Sample Type
  1. Rosmarinic acid attenuates hepatic fibrogenesis via suppression of hepatic stellate cell activation/proliferation and induction of apoptosis
    Authors: NM El-Lakkany, WH El-Maadawy, SH Seif El-Di, OA Hammam, SH Mohamed, SM Ezzat, MM Safar, S Saleh
    Asian Pac J Trop Med, 2017;10(5):444-453.
    Species: Rat
    Sample Type: Tissue Homogenates
  2. TIMP-1 inhibits the proteolytic processing of Reelin in experimental epilepsy.
    Authors: Tinnes, Stefanie, Ringwald, Julia, Haas, Carola A
    FASEB J, 2013;27(7):2542-52.
    Species: Rat
    Sample Type: Tissue Homogenates
  3. Antifibrotic effects of a recombinant adeno-associated virus carrying small interfering RNA targeting TIMP-1 in rat liver fibrosis.
    Authors: Cong M, Liu T, Wang P, Fan X, Yang A, Bai Y, Peng Z, Wu P, Tong X, Chen J, Li H, Cong R, Tang S, Wang B, Jia J, You H
    Am J Pathol, 2013;182(5):1607-16.
    Species: Rat
    Sample Type: Serum
  4. Nitric oxide induces TIMP-1 expression by activating the transforming growth factor beta-Smad signaling pathway.
    Authors: Akool el-S, Doller A, Muller R, Gutwein P, Xin C, Huwiler A, Pfeilschifter J, Eberhardt W
    J. Biol. Chem., 2005;280(47):39403-16.
    Species: Rat
    Sample Type: Cell Culture Supernates

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