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70-600 | Universal EZ-TFA Chemiluminescent Transcription Factor Assay

70-600
96 assays  
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      Replacement Information

      Tabela com principais espec.

      Species ReactivityKey ApplicationsDetection Methods
      M, H, R, Po, Eq, Rb, Gp, Ht, NhP, Ca ELISA, ACT Chemiluminescence
      Description
      Catalogue Number70-600
      Brand Family Upstate
      Trade Name
      • EZ-TFA
      • Upstate
      DescriptionUniversal EZ-TFA Chemiluminescent Transcription Factor Assay
      Materials Required but Not DeliveredReagents Cells, stimulated or treated as needed for the experimental system Capture probe Specific competitor Primary antibody Nuclear extraction kit (Catalog No. 2900) Nuclease free water (Catalog No. 3098) Equipment Heating Block Luminometer
      Background InformationThe UPSTATE Non-Radioactive Universal EZ-TFA Transcription Factor Assay provides a fast, sensitive method to detect specific transcription factor DNA binding activity in whole cell or nuclear extracts. This assay combines the principle of the electrophoretic mobility shift assay (EMSA) with the 96-well based enzyme-linked immunosorbent assay (ELISA), enabling manual or high-throughput sample analysis with greater sensitivity than conventional EMSA assays. After certain parameters are optimized, the entire assay takes less than 4 hours to complete with minimal hands-on time. The versatile set up allows for a flexible assay design and because the binding reaction occurs in solution, various parameters can be optimized such as probe titration, competitive oligonucleotide concentration, or treatment conditions. This kit contains the reagents and instructions necessary to develop an assay to monitor the DNA binding activity of any transcription factor of interest, or as a kit component for Upstate developed target specific Transcription Factor Assays. The Universal kit also allows for the design of native promoter capture probe sequences to interrogate specific transcription factor binding.

      Test Principle:
      The Non-Radioactive Universal Transcription Factor Assay kit is provided in a 96-well format. During the assay, the capture probe, a double stranded biotinylated oligonucleotide containing the consensus sequence for the specific transcription factor under study, is mixed with cellular (nuclear) extract in the transcription factor assay buffer provided directly in the streptavidin coated plate. When incubated together, the transcription factor contained in the extract binds to its consensus sequence. The biotinylated double stranded oligonucleotide bound to the transcription factor being studied is immobilized on the streptavidin-coated plate and any inactive, unbound material is washed away. The bound transcription factor is detected with a specific primary antibody. An HRP-conjugated secondary antibody is then used for detection and provides sensitive chemiluminescent detection that can be read in a microplate luminometer or by a CCD camera-coupled imaging system.
      References
      Product Information
      Components
      • Streptavidin Coated Chemiluminescent Plate, Catalog # 90494.
      • Blocking Reagent, Catalog # 90273.
      • 10X Annealing Buffer, Catalog # 2007044
      • Chemiluminescent Detection Reagent, Catalog #90495
      • Chemiluminescent Reaction Buffer, Catalog #90496
      • TFA Buffer, 5X, Catalog # 90272
      • Gt X Rb, HRP Secondary Antibody, Catalog # 90276
      • Gt X Ms, HRP Secondary Antibody, Catalog # 2003482
      Detection methodChemiluminescence
      Applications
      ApplicationThe Non-Radioactive Universal EZ-TFA Transcription Factor Assay provides a fast, sensitive method to detect specific transcription factor DNA binding activity in whole cell or nuclear extracts.
      Key Applications
      • ELISA
      • Activity Assay
      Biological Information
      Species Reactivity
      • Mouse
      • Human
      • Rat
      • Pig
      • Horse
      • Rabbit
      • Guinea Pig
      • Hamster
      • Non-human Primate
      • Canine
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated with gene specific target component with all necessary controls.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsUpon receipt, store components at the temperatures indicated on the labels.
      Store at 4°C:
      Transcription Factor Assay Chemiluminescent Plate, Catalog # 90494.
      Blocking Reagent, Catalog # 90273.
      10X Annealing Buffer, Catalog # 2007044.
      Chemiluminescent Detection Reagent, Catalog #90495.
      Chemiluminescent Reaction Buffer, Catalog #90496.
      Store at -20°C:
      Gt X Rb, HRP Secondary Antibody, Catalog # 90276.
      TFA Buffer, 5X, Catalog # 90272
      Gt X Ms, HRP Secondary Antibody, Catalog # 2003482.
      Packaging Information
      Material Size96 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Título

      Ficha de Segurança de Produtos (MSDS) 

      Certificados de análise

      TítuloNúmero do lote
      Universal EZ-TFA Chemiluminescent Transcription Factor Assay - manual manual

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      Anotai Atawong,Morifumi Hasegawa,Osamu Kodama
      Bioscience, biotechnology, and biochemistry 66 2002

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      Potassium ion efflux induced by cationic compounds in yeast.
      E Enríquez-Freire,R López,A Peña
      Biochimica et biophysica acta 1418 1999

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      GABA-dopamine receptor-receptor interactions in neostriatal membranes of the rat.
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      Neurochemical research 22 1997

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      Redox sites of NMDA receptors can modulate epileptiform activity in hippocampal slices from kainic acid-treated rats.
      O Quesada,J Hirsch,Y Ben-Ari,C Bernard
      Neuroscience letters 212 1996

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      Cholecystokinin-8 increases K(+)-evoked [3H] gamma-aminobutyric acid release in slices from various brain areas.
      M Pérez de la Mora,A M Hernandez-Gómez,J Méndez-Franco,K Fuxe
      European journal of pharmacology 250 1993

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      I Camacho-Arroyo,R Tapia
      Neurochemical research 17 1992

      Mostrar Resumo
      1356245 1356245
      Release of acetylcholine and GABA, and activity of their synthesizing enzymes in the rat pontine reticular formation.
      I Camacho-Arroyo,R Alvarado,R Tapia
      Neurochemical research 16 1991

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      Taurine release associated to volume regulation in rabbit lymphocytes.
      J Jesu? García,R Sánchez Olea,H Pasantes-Morales
      Journal of cellular biochemistry 45 1991

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      Journal of bioenergetics and biomembranes 23 1991

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      Brochura

      Título
      Shaping Epigenetics Discovery - Epigenetics Product Selection Brochure

      Informações técnicas

      Título
      96 well Template 2 Up
      96 well Template Single

      Manuais do usuário

      Título
      Universal EZ-TFA Transcription Factor Assay Chemiluminescent