ab181421 TNF alpha Human SimpleStep ELISA Ki...

Product nameTNF alpha Human SimpleStep ELISA? Kit See all TNF alpha kitsDetection methodColorimetricPrecision Intra-assay Sample n Mean SD CV% PBMC media 8 2.3% Inter-assay Sample n Mean SD CV% PBMC media 3 5% Sample typeSerum, Plasma, Cell culture mediaAssay typeSandwich (quantitative)Sensitivity 14 pg/mlRange31.25 pg/ml - 2000 pg/mlRecovery Sample specific recovery Sample type Average % Range Serum 99.5 80% - 119% Cell culture media 100.97 95% - 109% Heparin Plasma 98 91% - 106% EDTA Plasma 102 95.7% - 110% Citrate Plasma 93.09 83.5% - 99% Assay time1h 30m Assay durationOne step assaySpecies reactivityReacts with: Human Does not react with MouseProduct overview Abcam’s TNF-alpha in vitro SimpleStep ELISA? (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TNF-alpha protein in Human serum, plasma and culture media. 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 TNF-alpha, also known as cachectin or TNFSF1A, is the prototypic ligand of the TNF superfamily which plays a central role in inflammation, apoptosis, proliferation, invasion, angiogenesis, metastasis and morphogenesis. It is expressed on macrophages, endothelial, epithelial and tumor cells as a 26kDa transmembrane protein. TNF-alpha is cleaved by proteolytic processing into six chains: (1) TNF membrane form, (2) Intracellular domain 1, (3) Intracellular domain 2, (4) C-domain 1, (5) C-domain 2 and (6) TNF soluble form. Signaling from TNF-alpha differs depending on the type of ligand initiating the signaling event (intracellular, membrane or soluble). As an example, the membrane form of TNF-alpha appears to mediate anti-tumorigenic therapeutic responses whereas the soluble ligand is linked to inflammation and proliferation. Tested applicationsSandwich ELISAmore details PlatformMicroplate Properties Storage instructionsStore at +4°C. Please refer to protocols. Components 1 x 96 tests 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 5BI 2 x 3ml Plate Seals 1 unit Sample Diluent 50BP 2 x 12ml 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 TNFa Capture Antibody (Lyophilized) 2 vials TNFa Detector Antibody (Lyophilized) 2 vials TNFa Human Lyophilized Recombinant Protein 2 vials Research Areas Immunology Innate Immunity Cytokines Necrosis Factors Immunology Innate Immunity Cytokines TNF Superfamily Signal Transduction Growth Factors/Hormones TNF Stem Cells Signaling Pathways TGF beta TNF alpha Cancer Growth factors TNF Cancer Tumor immunology Cytokines TNF Kits/ Lysates/ Other Kits ELISA Kits ELISA Kits TNF ELISA kits FunctionCytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation.Involvement in diseaseGenetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).Sequence similaritiesBelongs to the tumor necrosis factor family.Post-translational modificationsThe soluble form derives from the membrane form by proteolytic processing. The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1. O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.Cellular localizationSecreted and Cell membrane. Information by UniProt Alternative names APC1 protein Cachectin DIF see allDatabase links Entrez Gene: 7124 Human Omim: 191160 Human SwissProt: P01375 Human Unigene: 241570 Human Applications Our Abpromise guarantee covers the use of ab181421 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.