Human TIMP-4 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DTM400
Control Products Available
Human TIMP-4 ELISA Calibrator Diluent RD6-31 Standard Curve
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Product Details
Procedure
Citations (5)
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Human TIMP-4 Quantikine ELISA Kit Summary

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 (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Human Milk (10 uL)
Sensitivity
10 pg/mL
Assay Range
78.1 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Human Milk)
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.

Product Summary

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.

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 252 793 1476 232 708 1377
Standard Deviation 11 46.9 77.8 20.8 45.8 96.4
CV% 4.4 5.9 5.3 9 6.5 7

Serum, EDTA Plasma, Heparin Plasma, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 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

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.
Human TIMP-4 ELISA Linearity

Data Examples

Human TIMP-4 ELISA Calibrator Diluent RD6-31 Standard Curve

Human TIMP-4 ELISA Calibrator Diluent RD6-31 (1X) Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TIMP-4

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 4
Entrez Gene IDs:
7079 (Human); 110595 (Mouse)
Alternate Names:
metalloproteinase inhibitor 4; TIMP metallopeptidase inhibitor 4; TIMP4; TIMP-4; tissue inhibitor of metalloproteinase 4; Tissue inhibitor of metalloproteinases 4
⚠ 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.

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. 100 µL Assay Diluent
  4.   Add 100 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 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

Citations for Human TIMP-4 Quantikine ELISA Kit

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.

5 Citations: Showing 1 - 5
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  1. Circulating biomarkers of tissue remodelling in pulmonary hypertension.
    Authors: Schumann C, Lepper PM, Frank H, Schneiderbauer R, Wibmer T, Kropf C, Stoiber KM, Rudiger S, Kruska L, Krahn T, Kramer F
    Biomarkers, 2010;15(0):523.
    Species: Human
    Sample Types: Plasma
  2. Profile of macrophages in human abdominal aortic aneurysms: a transcriptomic, proteomic, and antibody protein array study.
    Authors: Lamblin N, Ratajczak P, Hot D, Dubois E, Chwastyniak M, Beseme O, Drobecq H, Lemoine Y, Koussa M, Amouyel P, Pinet F
    J. Proteome Res., 2010;9(7):3720-9.
    Species: Human
    Sample Types: Plasma
  3. Primary human acute myelogenous leukemia cells release matrix metalloproteases and their inhibitors: release profile and pharmacological modulation.
    Authors: Reikvam H, Hatfield KJ, Oyan AM, Kalland KH, Kittang AO, Bruserud O
    Eur. J. Haematol., 2010;84(3):239-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Imbalance of tissue inhibitors of metalloproteinases (TIMP) - 1 and - 4 serum levels, in patients with inflammatory bowel disease.
    Authors: Kapsoritakis AN, Kapsoritaki AI, Davidi IP, Lotis VD, Manolakis AC, Mylonis PI, Theodoridou AT, Germenis AE, Potamianos SP
    BMC Gastroenterol, 2008;8(1):55.
    Species: Human
    Sample Types: Serum
  5. Specific temporal profile of matrix metalloproteinase release occurs in patients after myocardial infarction: relation to left ventricular remodeling.
    Authors: Webb CS, Bonnema DD, Ahmed SH, Leonardi AH, McClure CD, Clark LL, Stroud RE, Corn WC, Finklea L, Zile MR, Spinale FG
    Circulation, 2006;114(10):1020-7.
    Species: Human
    Sample Types: Plasma

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