Human MMP-9 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MMP-9 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-9 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for MMP-9 is added to the wells and binds to the combination of capture antibody- MMP-9 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of MMP-9 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven MMP-9 standard dilutions and MMP-9 sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade extracellular matrix proteins. They are secreted as zymogens (Pro-MMPs) that are activated by a variety of proteinases or by reaction with organic mercurials. They are inhibited by specific tissue inhibitors of metalloproteinases (TIMPs) and by 2-macroglobulin. The regulation of MMP activity is important in tissue remodeling, inflammation, tumor growth and metastasis.

Human MMP-9 (also known as gelatinase B) is secreted as a 92 kDa zymogen. Cleavage of Pro-MMP-9 at or near residue 87 results in the active enzyme with a mass of approximately 82 kDa. MMP-9 has three fibronectin type II domains, a hemopexin-like domain and a proline-rich type V collagen-homologous domain. Pro-MMP-9 can be activated by MMP-3 or by certain bacterial proteinases. MMP-9 is inhibited by α2-macroglobulin or by TIMP-1, which binds to Pro-MMP-9 as well as to active MMP-9. In vitro treatment of Pro-MMP-9 with 4-aminophenylmercuric acid (APMA) produces not only the 82 kDa active enzyme but also a C-terminal truncated form of approximately 65 kDa with the activity comparable to that of the 82 kDa form.

Pro-MMP-9 is secreted by monocytes, macrophages, neutrophils, keratinocytes, fibroblasts, osteoclasts, chondrocytes, skeletal muscle satellite cells, endothelial cells, and various tumor cells. Pro-MMP-9 expression is upregulated by TGF-β1, IL-1β, TGF-α, PDGF-AB, TNF-α, and IL-1α. Substrates for MMP-9 include denatured collagen type I (gelatin), native collagens type IV, V, VII, X and XI, fibrinogen, vitronectin, IL-1β, and entactin, a molecule that bridges laminin and type IV collagen.

GENE ID	4318
SWISS PROT	P14780
SYNONYMS	GELB; CLG4B; MMP-9; MANDP2

Materials Supplied

Kit Components	96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human MMP-9 (8﹡12)	1 plate
Human MMP-9 Standard lyophilized, 4000 pg/ml upon reconstitution	2 vials
Concentrated Biotin-Conjugate anti-human MMP-9 monoclonal antibody	2 vials
Streptavidin-HRP solution	2 vials
Standard /sample Diluent	1 bottle
Biotin-Conjugate antibody Diluent	1 bottle
Streptavidin-HRP Diluent	1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20)	1 bottle
Substrate Solution	1 vial
Stop Solution	1 vial
Adhesive Films	4 pieces
Product data sheet	1 copy

Storage

Store at 2 - 8°C

Performance Characteristics

REPEATABILITY	The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY	The minimum detectable dose was 30pg/mL.
ASSAY RANGE	62.50 - 4000 pg/mL
SPECIFICITY	This assay recognizes both natural and recombinant human MMP-9. The factors listed below were prepared at 200ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.

Factors assayed for cross-reactivity

Recombinant human	MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-10, MMP-12
Recombinant mouse	MMP-2, MMP-3, MMP-9, TIMP-1

Data Analysis Assistance

We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKH1042.

https://www.myassays.com/bioss-human-mmp-9-elisa-kit.assay