Human Pro-MMP-1 Quantikine ELISA Kit

R&D Systems | Catalog # DMP100

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), Heparin Plasma (100 µL)

Sensitivity

0.095 ng/mL

Assay Range

0.156-10 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma)
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Product Summary for Human Pro-MMP-1 Quantikine ELISA Kit

The Quantikine Human Pro-MMP-1 Immunoassay is a 4.5 hour solid phase ELISA designed to measure Pro-MMP-1 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Pro-MMP-1 and antibodies raised against the recombinant factor. The antibodies will not recognize recombinant human active MMP-1 or TIMP bound MMP-1. Natural human Pro-MMP-1 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 mass values of natural human Pro-MMP-1.

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 Pro-MMP-1

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 (ng/mL) 0.494 1.65 3.39 0.561 1.73 3.50
Standard Deviation 0.022 0.065 0.177 0.065 0.153 0.270
CV% 4.5 4.0 5.2 11.7 8.8 7.7

Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 0.702 2.13 4.43 0.738 2.34 4.66
Standard Deviation 0.039 0.130 0.241 0.077 0.176 0.312
CV% 5.6 6.1 5.4 10.4 7.5 6.7

Recovery for Human Pro-MMP-1 Quantikine ELISA Kit

The recovery of Pro-MMP-1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 98-112
Heparin Plasma (n=5) 92 85-102
Serum (n=5) 97 87-112

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Pro-MMP-1 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.\line

Human Pro-MMP-1 ELISA Linearity

Scientific Data Images for Human Pro-MMP-1 Quantikine ELISA Kit

Human Pro-MMP-1 ELISA Cell Culture Supernate Standard Curve

Human Pro-MMP-1 ELISA Cell Culture Supernate Standard Curve

Human Pro-MMP-1 ELISA Serum/Plasma Standard Curve

Human Pro-MMP-1 ELISA Serum/Plasma 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: MMP-1

The matrix metalloproteinases (MMPs) consist of 24 known human zinc proteases with essential roles in breaking down components of the extracellular matrix (ECM). Additional MMP substrates include cytokines, chemokines, growth factors and binding proteins, cell/cell adhesion molecules, and other proteinases. With a few exceptions, MMPs share common structural motifs including a pro-peptide domain, a catalytic domain, a hinge region, and a hemopexin-like domain. Synthesized as pro-enzymes, most MMPs are secreted before conversion to their active form. MMP activities are modulated on several levels including transcription, pro-enzyme activation, or by their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs). A subset of MMPs are associated with membranes and designated as membrane-type metalloproteinases (MT-MMP).

Long Name

Matrix Metalloproteinase 1

Alternate Names

MMP1

Entrez Gene IDs

4312 (Human); 83995 (Mouse)

Gene Symbol

MMP1

Additional MMP-1 Products

Product Documents for Human Pro-MMP-1 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 Pro-MMP-1 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.

Citations for Human Pro-MMP-1 Quantikine ELISA Kit

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    Sample Tested: Serum-free Cell Culture Media
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Protocols

View specific protocols for Human Pro-MMP-1 Quantikine ELISA Kit (DMP100):

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. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  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 on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well.
  13.   For Cell Culture Supernate Samples: Incubate at room temperature for 20 minutes on the benchtop. PROTECT FROM LIGHT.
    For Serum & Plasma Samples: Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  14. 50 µL Stop Solution
  15. 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.

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