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RayBiotech/Custom Human Phospho-C-Fos (Thr232) and Total C-Fos Cell-Based ELISA/1 Plate Kit/CBEL-CFos-T232-1
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Antigen InformationAccession Number:P01100Gene ID:2353Gene Symbols:FOSG0S7Protein Name & Synonyms:Proto-oncogene c-Fos (Cellular oncogene fos) (G0/G1 switch regulatory protein 7)Target Species:HumanAssay FormatPathway:IGF SignalingJAK/STAT SignalingMAPK SignalingPI3K-AKT SignalingPKC SignalingTGF-beta SignalingSpecificity:This ELISA kit recognizes Human C-Fos phosphorylated at site Threonine-232 as well as total C-Fos.Number of Targets Detected:2Species Detected:HumanCompatible Sample Types:Adherent Cell CultureDesign Principle:Cell-basedMethod of Detection:ColorimetricQuantitative/Semi-Quantitative:Semi-QuantitativeSolid Support:96-well MicroplateProduct SpecificationsSize:1, 2, or 5 x 96-Well Strip Microplate KitProduct FeaturesRapidly measure phosphorylated protein in adherent cell linesSimultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)No sample lysis is neededCompatible with a standard ELISA plate readerApplication NotesKit ComponentsUncoated 96-well Strip MicroplateWash BuffersFixing SolutionQuenching BufferBlocking BufferAnti-phospho antibodyAnti-pan antibodyHRP-Conjugated Secondary AntibodyTMB One-Step SubstrateStop SolutionOther Materials RequiredDistilled or deionized water100 ml and 1 liter graduated cylindersTubes to prepare sample dilutionsProtease and Phosphatase inhibitorsPrecision pipettes to deliver 2 µl to 1 ml volumesAdjustable 1-25 ml pipettes for reagent preparationBenchtop rocker or shakerMicroplate reader capable of measuring absorbance at 450 nmProtocol OutlineSeed 10,000-30,000 cells into each well and incubate overnight.Apply various treatment, inhibitors or activators according to manufacturer"s instructions.Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.Add 200 µl of Blocking Solution and incubate for 1 h at 37°C.Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.Add 100 µl of TMB One-Step Substrate Reagent to each well.Incubate 30 min at RT.Add 50 µl of Stop Solution to each well.Read at 450 nm immediately.Storage/StabilityUpon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.No posts foundWrite A ReviewManualSDS

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