Thermo Scientific™ LightShift™ EMSA Optimization and Control Kit

Thermo Scientific™ LightShift™ EMSA Optimization and Control Kit

The Thermo Scientific LightShift EMSA Optimization and Control 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 and a control set of DNA and protein extract to test the kit system.
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
• For Use With (Equipment): myECL™ Imager, X-Ray Film
• Assay: EMSA Assay
• Product Type: LightShift EMSA Optimization and Control Kit
• Target Specificity: Not Target-Specific
• Technique: Gel Shift

Contents

• 10X Binding Buffer, 1 mL
• Biotin-EBNA Control DNA, 50 µL
• Unlabeled EBNA DNA, 50 µL
• EBNA Extract, 125 µL
• Poly (dI·dC), 125 µL
• 50% Glycerol, 500 µL
• 1% NP-40, 500 µL
• 1 M KCl, 1 mL
• 100mM MgCl2, 500 µL
• 200mM EDTA pH 8.0, 500 µL
• 5X Loading Buffer, 1 mL