Mouse MMP-3 (Matrix Metalloproteinase 3) ELISA Kit (MOES01134)

SKU:
MOES01134
€549

Description

ELISA Kit Technical ManualMSDS

Mouse MMP-3 (Matrix Metalloproteinase 3) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse MMP-3 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse MMP-3 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse MMP-3, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse MMP-3. The concentration of Mouse MMP-3 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection MethodColormetric
Detection Range0.16-10 ng/mL
Sensitivity0.10 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse MMP-3 in samples. No significant cross-reactivity or interference between Mouse MMP-3 and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(ng/mL)
O.D Average Corrected
102.394
2.412
2.4032.318
51.468
1.518
1.4931.408
2.50.858
0.856
0.8570.772
1.250.436
0.47
0.4530.368
0.630.245
0.243
0.2440.159
0.310.179
0.175
0.1770.092
0.160.128
0.14
0.1340.049
00.076
0.094
0.085--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse MMP-3 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse MMP-3 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.501.104.200.501.204.60
Standard deviation0.030.060.210.030.060.17
C V (%)6.005.455.006.005.003.70

Recovery

The recovery of Mouse MMP-3 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)92-10697
EDTA plasma (n=5)87-10394
Cell culture media (n=5)95-108100

Linearity

Samples were spiked with high concentrations of Mouse MMP-3 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)88-10292-10886-100
Average (%)949891
1:4Range (%)99-11483-9695-107
Average (%)10588101
1:8Range (%)96-10687-9993-105
Average (%)1019298
1:16Range (%)96-11383-9496-113
Average (%)10489103

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse MMP-3 (Matrix Metalloproteinase 3) ELISA Kit (MOES01134) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Mouse MMP-3 (Matrix Metalloproteinase 3) ELISA Kit (MOES01134) Protein Information

UniProt Protein Function:MMP3: Can degrade fibronectin, laminin, gelatins of type I, III, IV, and V; collagens III, IV, X, and IX, and cartilage proteoglycans. Activates procollagenase. Defects in MMP3 are the cause of susceptibility to coronary heart disease type 6 (CHDS6). A multifactorial disease characterized by an imbalance between myocardial functional requirements and the capacity of the coronary vessels to supply sufficient blood flow. Decreased capacity of the coronary vessels is often associated with thickening and loss of elasticity of the coronary arteries. A polymorphism in the MMP3 promoter region is associated with the risk of coronary heart disease and myocardial infarction, due to lower MMP3 proteolytic activity and higher extracellular matrix deposition in atherosclerotic lesions. Belongs to the peptidase M10A family.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Secreted; Protease; Secreted, signal peptide; EC 3.4.24.17

Cellular Component: extracellular matrix; proteinaceous extracellular matrix; extracellular space; protein complex; mitochondrion; dendrite; extracellular region; cytosol; nucleus

Molecular Function:peptidase activity; hydrolase activity; metallopeptidase activity; zinc ion binding; metalloendopeptidase activity; metal ion binding; endopeptidase activity; protein complex binding; calcium ion binding

Biological Process: positive regulation of protein oligomerization; collagen catabolic process; negative regulation of protein kinase B signaling cascade; protein catabolic process; proteolysis; regulation of cell migration; positive regulation of cell migration

NCBI Summary:This gene encodes a member of the matrix metalloproteinase family of extracellular matrix-degrading enzymes that is involved in tissue remodeling, wound repair, progression of atherosclerosis and tumor invasion. The encoded protein is activated by the removal of an N-temrinal activation peptide to generate a zinc-dependent endopeptidase with a broad range of substrates such as proteoglycans, laminin, fibronectin, elastin, and collagens. This gene is located in a cluster of other matrix metalloproteinase genes on chromosome 9. [provided by RefSeq, Aug 2015]
UniProt Code:P28862
NCBI GenInfo Identifier:461769
NCBI Gene ID:17392
NCBI Accession:P28862.2
UniProt Related Accession:P28862
Molecular Weight:53,845 Da
NCBI Full Name:Stromelysin-1
NCBI Synonym Full Names:matrix metallopeptidase 3
NCBI Official Symbol:Mmp3  
NCBI Official Synonym Symbols:SLN1; Str1; SLN-1; STR-1; Stmy1  
NCBI Protein Information:stromelysin-1; SL-1; EMS-2; MMP-3; transin-1; progelatinase; stromelysin 1; matrix metalloproteinase 3; matrix metalloproteinase-3
UniProt Protein Name:Stromelysin-1
UniProt Synonym Protein Names:EMS-2; Matrix metalloproteinase-3; MMP-3; Transin-1
Protein Family:Stromelysin
UniProt Gene Name:Mmp3  
UniProt Entry Name:MMP3_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
Sandwich
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