FTO (mouse intracellular) ELISA Kit

FTO, Fat mass-and obesity-associated gene, was discovered as a responsible gene causing the mouse ‘fused toes’ mutation. The predicted 502-amino acid Fto protein has a calculated molecular mass of 58 kD and contains an N-terminal bipartite nuclear localization signal. RT-PCR detected Fto expression throughout mouse embryonic development and in all adult mouse tissues examined except heart and skin. Bioinformatics analysis indicates that FTO shares sequence motifs with iron- and 2-oxoglutarate (2OG)-dependent oxygenases. Recombinant murine Fto catalyzes the iron- and 2OG-dependent demethylation of 3-methylthymine in single-stranded DNA, with concomitant production of succinate, formaldehyde, and carbon dioxide. In wildtype mice, Fto mRNA is most abundant in the brain, particularly in hypothalamic nuclei, and are regulated by feeding and fasting. This assay is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for quantitative determination of mouse FTO in cells. A monoclonal antibody specific for FTO has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, FTO is recognized by the addition of a purified polyclonal antibody specific for FTO (Detection Antibody). After removal of excess polyclonal antibody, HRP conjugated anti-rabbit IgG (Detector) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450 nm after acidification and is directly proportional to the concentration of FTO in the samples. This ELISA is specific for the measurement of natural and recombinant mouse FTO. It does cross-react with mouse adiponectin, mouse RBP4, mouse Nampt, mouse vaspin, mouse progranulin, mouse resistin, mouse clusterin, mouse GPX3, mouse CD137, rat adiponectin, rat Nampt, human adiponectin, human Nampt, human progranulin. The assay range is 0.156 – 10 ng FTO/ml and a detection limit of 20 pg/ml (based on adding two standard deviations to the mean value of the (50) zero standards).

• Detection method: Absorbance (450 nm) • Species reactivity: Mammalian • Applications: This assay is to be used for the in vitro quantitative determination of mouse FTO in the range of 0.156 – 10 ng FTO/ml and a detection limit of 20 pg/ml (based on adding two standard deviations to the mean value of the (50) zero standards).

• Simple procedure • Fast and convenient • High-throughput adaptable

FTO

none

• Pre-coated Microtiter Plate • Wash Buffer (10X) • Diluent (5X) • Lysis Buffer (10X) • Detection Antibody • Detector 100X (Hrp conjugated anti-IgG) • Mouse FTO Standard (lyophilized, 20 ng) • Mouse FTO QC Sample (lyophilized) • TMB Substrate Solution • Stop Solution • Plate Sealers

+4°C

gel pack

12 months

see datasheet

Inquire

Cell lysates

E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays,E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays

ELISA Enzyme-linked immunosorbent assays Code 90320007 SNOMED,Mouse or mice from the Mus musculus species are used for production of mouse monoclonal antibodies or mabs and as research model for humans in your lab. Mouse are mature after 40 days for females and 55 days for males. The female mice are pregnant only 20 days and can give birth to 10 litters of 6-8 mice a year. Transgenic, knock-out, congenic and inbread strains are known for C57BL/6, A/J, BALB/c, SCID while the CD-1 is outbred as strain.

Mus musculus