Human MMP14 ELISA Kit (ab197747)
- Datasheet
- References (1)
- Protocols
Overview
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Product name
Human MMP14 ELISA Kit
See all MMP14 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Overall 8 2.94% Inter-assay Sample n Mean SD CV% Overall 3 2.52% -
Sample type
Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
0.137 ng/ml -
Range
0.391 ng/ml - 25 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 95 93% - 98% Cell culture media 83 79% - 88% Hep Plasma 90 88% - 92% EDTA Plasma 91 90% - 92% Cit plasma 94 91% - 100% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
MMP14 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MMP14 protein in human serum, plasma, cell culture supernatants, cell and tissue extracts.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Sensitivity:
Samples diluted in Sample Diluent NS – 0.145 ng/mL
Samples diluted in 1X Cell Extraction Buffer – 0.137 ng/mL -
Notes
MMP14 is one of six membrane type matrix metalloproteinases, and was the first membrane type MMP discovered. Matrix metalloproteinases are zinc binding enzymes that degrade components of the extracellular matrix, and are involved in tissue remodeling, wound healing, angiogenesis, and tumor invasion. MMP14 may specifically activate progelatinase A and, through pro-gelatinase A activation, trigger tumor cell invasion on the tumor cell surface. MMP14 may also be involved in actin cytoskeleton reorganization by cleaving PTK7, as well as be a positive regulator of cell growth and migration through the activation of MMP15. In mice that were MMP14 gene deficient dysmorphism, arthritis, osteopenia, dwarfism, and fibrosis were observed, which are all conditions related to skeletal and soft tissue modeling.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human MMP14 Capture Antibody 1 x 600µl 10X Human MMP14 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 1 x 6ml Human MMP14 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Function
Seems to specifically activate progelatinase A. May thus trigger invasion by tumor cells by activating progelatinase A on the tumor cell surface. May be involved in actin cytoskeleton reorganization by cleaving PTK7. Acts as a positive regulator of cell growth and migration via activation of MMP15. -
Tissue specificity
Expressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors. -
Sequence similarities
Belongs to the peptidase M10A family.
Contains 4 hemopexin-like domains. -
Domain
The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme. -
Post-translational
modificationsThe precursor is cleaved by a furin endopeptidase. -
Cellular localization
Membrane. Melanosome. Cytoplasm. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Forms a complex with BST2 and localizes to the cytoplasm. - Information by UniProt
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Alternative names
- Matrix metalloproteinase-14
- Membrane type 1 metalloprotease
- Membrane-type matrix metalloproteinase 1
see all -
Database links
- Entrez Gene: 4323 Human
- Omim: 600754 Human
- SwissProt: P50281 Human
- Unigene: 2399 Human
Applications
Our Abpromise guarantee covers the use of ab197747 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Other - Human MMP14 ELISA Kit (ab197747)SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of MMP14 standard curve prepared in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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Example of MMP14 standard curve prepared in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Titration of Human THP-1 cell extract and Human liver homogenate tissue extract within the working range of the assay.The interpolated MMP14 concentration of each lysate dilution is shown.
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Titration of Human serum, plasma, and cell media (+ 10% FBS) spiked with concentrations of recombinant MMP14 protein within the working range of the assay.Background-subtracted data values are graphed.
Protocols
References
This product has been referenced in:
- Schmitt R et al. A potential key mechanism in ascending aortic aneurysm development: Detection of a linear relationship between MMP-14/TIMP-2 ratio and active MMP-2. PLoS One 14:e0212859 (2019). Read more (PubMed: 30794673) »
