ab181419 Fibronectin Human SimpleStep ELISA ...

Product nameFibronectin Human SimpleStep ELISA? Kit See all Fibronectin kitsDetection methodColorimetricPrecision Intra-assay Sample n Mean SD CV% HeLa extract 5 7.3% Inter-assay Sample n Mean SD CV% HeLa extract 3 9% Sample typeCell culture supernatant, Urine, Serum, PlasmaAssay typeSandwich (quantitative)Sensitivity 70 pg/mlRange200 pg/ml - 12500 pg/mlRecovery Sample specific recovery Sample type Average % Range Urine 112 111% - 114% Serum 97 91% - 109% Cell culture media 86 85% - 86% EDTA Plasma 92 88% - 103% Citrate Plasma 104 95% - 116% Assay time1h 30m Assay durationOne step assaySpecies reactivityReacts with: Human Product overview Abcam’s Fibronectin in vitro SimpleStep ELISA? (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Fibronectin protein in Human serum, plasma, cell culture supernatant and urine. 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. Notes Fibronectin (FN) is a large glycoprotein (220 kDa) present in the extracellular matrix and circulating plasma. Fibronectin is important in many cell adhesion and migration related processes, including wound healing, embryogenesis and nerve regeneration. Differential expression of fibronectin is seen in coronary heart disease, glomerulopathy and tumor cell metastasis. The protein contains binding sites for collagen, heparin and fibrin and is a specific ligand for several integrin adhesion receptors. Fibronectin exists as a dimer or multimer and is encoded by the FN1 gene. Tested applicationsSandwich ELISAmore details PlatformMicroplate Properties Storage instructionsStore at +4°C. Please refer to protocols. Components 1 x 96 tests 10X Fibronectin Capture Antibody 2 x 300μl 10X Fibronectin Detector Antibody 2 x 300μl 10X Wash Buffer PT 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 2 x 3ml Fibronectin Human Lyophilized Purified Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate 1 unit Stop Solution 1 x 12ml TMB Substrate 1 x 12ml Research Areas Signal Transduction Cytoskeleton / ECM Extracellular Matrix ECM Proteins Fibronectin Cancer Invasion/microenvironment ECM Extracellular matrix Other Kits/ Lysates/ Other Kits ELISA Kits ELISA Kits Adhesion molecules ELISA kits FunctionFibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization. Participates in the regulation of type I collagen deposition by osteoblasts. Anaslin binds fibronectin and induces fibril formation. This fibronectin polymer, named superfibronectin, exhibits enhanced adhesive properties. Both anaslin and superfibronectin inhibit tumor growth, angiogenesis and metastasis. Anaslin activates p38 MAPK and inhibits lysophospholipid signaling.Tissue specificityPlasma FN (soluble dimeric form) is secreted by hepatocytes. Cellular FN (dimeric or cross-linked multimeric forms), made by fibroblasts, epithelial and other cell types, is deposited as fibrils in the extracellular matrix. Ugl-Y1, Ugl-Y2 and Ugl-Y3 are found in urine.Involvement in diseaseGlomerulopathy with fibronectin deposits 2Sequence similaritiesContains 12 fibronectin type-I domains. Contains 2 fibronectin type-II domains. Contains 16 fibronectin type-III domains.Developmental stageUgl-Y1, Ugl-Y2 and Ugl-Y3 are present in the urine from 0 to 17 years of age.Post-translational modificationsSulfated. It is not known whether both or only one of Thr-2064 and Thr-2065 are/is glycosylated. Forms covalent cross-links mediated by a transglutaminase, such as F13A or TGM2, between a glutamine and the epsilon-amino group of a lysine residue, forming homopolymers and heteropolymers (e.g. fibrinogen-fibronectin, collagen-fibronectin heteropolymers). Phosphorylation sites are present in the extracellular medium. Proteolytic processing produces the C-terminal NC1 peptide, anaslin.Cellular localizationSecreted > extracellular space > extracellular matrix. Information by UniProt Alternative names CIG Cold insoluble globulin Cold-insoluble globulin see allDatabase links Entrez Gene: 2335 Human Omim: 135600 Human SwissProt: P02751 Human Unigene: 203717 Human Applications Our Abpromise guarantee covers the use of ab181419 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.