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Gel Shift Assay Systems

Simple, rapid systems for detecting DNA-binding proteins

Manufacturer: promega corportaion  E3050

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Catalog No. PR-E3050

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Description & Specifications


Product Type Gel Shift Assay Core System
For Use With (Application) Used to study sequence specific DNA binding Proteins such as transcription facto rs
Quantity 100 Reactions
Includes HeLaScribe™ nuclear extract, 40μL Gel shift assay grade, T4 Polynucleotide Kinase, 100U Kinase Buffer (10X), 100μL Gel shift Binding Buffer (5X), 200μL, AP2 and SP1 Consensus Oligonucleotides, 35pmol each.
Storage Requirements HeLa nuclear extract at -70°C and other components at -20°C

The gel shift assay is based on the observation that complexes of protein and DNA migrate through a nondenaturing polyacrylamide gel more slowly than free DNA fragments or double-stranded oligonucleotides. The specificity of a DNA-binding protein for the suspected binding site is established by competition experiments using a 32P end-labeled DNA fragment containing the suspected binding site and unlabeled DNA fragments or oligonucleotides containing a binding site for the protein of interest or other unrelated DNA sequences.

  • The Core System contains an E. coli extract containing cloned AP2 protein and an AP2 consensus oligo, making it a reliable system for gaining experience with gel shift assays, due to the strong gel shift produced by AP2
  • The Complete System contains five additional double-stranded oligonucleotides that represent consensus sequences to characterized binding sites
  • Provided oligonucleotides can be 5« end-labeled and used as protein-specific probes or used as unlabeled oligonucleotides in competition assays