Human MMP-9 ELISA Kit

Detection Principle:

This kit uses double antibody sandwich enzyme-linked immunosorbent assay technology. The specific anti human mmp-9antibody was pre coated on a high affinity microplate. The standard, sample to be tested and biotinylated detection antibody are added into the wells of the enzyme plate. After incubation, the mmp-9present in the sample combines with the solid-phase antibody and detection antibody to form an immune complex. After washing to remove unbound material, horseradish peroxidase labeled streptavidin (streptavidin HRP) was added. After washing, the chromogenic substrate was added to avoid light for color development. Stop the reaction by adding stop solution, and determine the absorbance value at 450 nmwavelength (reference correction wavelength 540nmor 570nm).

Test Type: Double antibody sandwich method

Form: pre coated 96orifice plate

Test Sample Type: cell supernatant, Serum, plasma

Sample Loading: 100ul

Kit Components:

Pre coated 96well plates, standards, mmp-9detection antibody, dilution buffer, chromogenic solution (a, b)One copy of washing solution, termination solution, sa-hrp, plate sealing membrane and instructions.

Sensitivity: 0.156ng/ml

Detection Range: 31.2 - 2000 pg/ml

Recovery Range: 85-104%

Saving Method: 2-8℃

Standard Curve

Background:

Matrix metalloproteinases (MMPs), also known as matrixins, are members of the family of zinc calcium dependency proteolytic enzyme, can decompose the extracellular matrix (ECM), in different subcellular environment, to participate in a variety of molecular processes. MMPs play important functions in numerous physiological processes, such as embryonic development, morphogenesis, reproduction, and tissue remodeling. Also involved in inflammation and autoimmune diseases, such as arthritis, cancer and cardiovascular disease. The quantity of the new synthesis of MMPs main in transcription regulation, the already existing MMPs proteolytic activity, not only controlled by zymogenic activity, is also controlled by inhibition of endogenous inhibitor of enzyme activity, such as & alpha; 2-macroglobulin, and tissue inhibitors of metalloproteinases (TIMPs).

"MMP-9 (also known as gelatinase B, 92kDa type IV collagenase, 92kDa gelatinase, and type V collagenase) is secreted as a proglycosylase." Activation of the zymogens involves proteolytic removal of the N-terminal precursor region, resulting in the formation of the 82-kda active enzyme. Active human MMP-9 shares 72% and 74% amino acid sequence identity with mouse and rat MMP-9, respectively. In addition to the zinc binding sites, catalytic domain structure also contains three consecutive fibronectin II homologous unit, can be combined with gelatin. Hinge region rich in proline connect catalytic domain structure to C - end heme sample structure domain. In vitro, treatment of the proenzyme with 4-aminophenyl acetate (APMA) produced not only the active enzyme but also a truncated C-terminal form with comparable activity. MMP - 9 can degrade component of ECM, especially to the degeneration of collagen (gelatin) has a very high specificity. MMP-9 also cleaves collagen types III, IV, V, and XI, as well as elastin, nestin, and vitronectin. MMP-9 also cleases a variety of chemokines and growth factors (e.g., IL-1β , CXCL8 / IL - 8 CXCL7 CXCL4, CXCL1, Latent TGF - & beta; , the combination of membrane and TNF - & alpha; , VEGF and FGF basic), & beta; -Amyloid, substance P, myelin basic protein. "This action may increase or decrease the bioactivity of these soluble factors, and may also release them from association with the ECM." MMP - 9 can also be triggered by all kinds of membrane protein signaling pathways, or inducing they fell from the cell membrane to inhibit signaling pathways (such as CD44, E - calcium mucins, integrin, ICAM - 1 and IL - 2 Rα) .

MMP - 9 is produced by a variety of normal cells and the cells, including neutrophils, monocytes, macrophages, astrocytes, fibroblast, broken bone cells, cartilage cells, keratinocytes, endothelial cells and epithelial cells. It affects physiological and pathological angiogenesis and vascular remodeling. Activated neutrophils release MMP-9 precursors, which do not bind TIMP-1, allowing pro-angiogenic FGF-2 to be released from the ECM. MMP - 9 and TIMP - 1 compounds can induce the release of FGF - 2. Neutrophils sources of MMP - 9 can aggravate the inflammatory response, induced by collagen polypeptide produced from the release of extra neutrophils MMP - 9. MMP - 9 in bone formation and remodeling, induced behavioral sensitization and methamphetamine response, regulation and control of the reconstruction of the synapses, the trophoblast invasion in the process of implantation, serine protease inhibitors & alpha; 1, the activation of also play an important role. MMP-9 mediated shedding of adhesion molecules also plays a direct role in tumor cell invasion.

Circulating levels of MMP-9 are elevated in a number of inflammatory disorders, including luminal thrombosis, atherosclerosis, Crohn's disease, hepatitis C virus infection, colorectal cancer, and Duchenne muscular dystrophy. The proportion of MMP 9 and TIMP - 1 in multiple sclerosis in the serum and cystic fiber phlegm, but decrease in serum in cytomegalovirus infection. The levels of free MMP-9 and the complex of MMP-9 and lipocalin-2 /NGAL were increased in the urine of patients with ovarian cancer and patients with uterine and urinary tract infection, respectively.