Thermo Scientific™ LightShift™ Chemiluminescent EMSA Kit

Thermo Scientific™ LightShift™ Chemiluminescent EMSA Kit

The Thermo Scientific LightShift Chemiluminescent EMSA Kit is an extraordinarily robust and sensitive system for performing electrophoretic mobility shift assays (EMSA) to identify and characterize protein-DNA binding interactions. The kit includes reagents for setting up and customizing DNA binding reactions, a control set of DNA and protein extract to test the kit system, stabilized streptavidin-HRP conjugate to probe for the biotinlabeled DNA target, and an exceptionally sensitive chemiluminescent substrate module for detection.
The principle for LightShift EMSA Detection is similar to a Western blot. Biotin end-labeled duplex DNA is incubated with a nuclear extract or purified factor and electrophoresed on a native gel. The DNA is then rapidly (30 minutes) transferred to a positive nylon membrane, UV crosslinked, probed with streptavidin-HRP conjugate and incubated with the substrate. The protocol from labeling to results can be accomplished in a single day.

Features

• Excellent for detecting low-abundance proteins in nuclear extracts
• Sensitivity that surpasses radioactive and digoxigenin methods
• Compatible with previously established binding conditions for popular DNA-protein interactions
• Includes EBNA control system to help new users develop a working assay and understand the methods used to confirm binding interaction specificity
  

Contents

• 10X Binding Buffer: 1mL
• Biotin–EBNA Control DNA: 50μL
• Unlabeled EBNA DNA: 50μL
• Epstein-Barr Nuclear Antigen (EBNA) Extract: 125μL
• Poly (dI•dC): 125μL
• 50% Glycerol: 500μL
• 1% NP-40: 500μL
• 1 M KCl: 1mL
• 100mM MgCl2: 500μL
• 200mM EDTA pH 8.0: 500μL
• 5X Loading Buffer: 1mL
• Stabilized Streptavidin-HRP, 1.5 mL (store at 4°C)
• Luminol/Enhancer Solution, 80 mL (store at 4°C)
• Stable Peroxidase Solution, 80 mL (store at 4°C)
• Blocking Buffer, 500 mL (store at 4°C)
• 4X Wash Buffer, 500 mL (store at 4°C)
• Substrate Equilibration Buffer, 500 mL (store at 4°C)