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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 mobilityRead more
Catalog number 20148X
Price (USD)
235.00
Each
Price (USD)
235.00
Each
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.
Features of theLightShift EMSA Optimization and Control Kit:
• 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
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.
The interaction of proteins with DNA is central to the control of many cellular processes including DNA replication, recombination and repair, transcription, and viral assembly. One technique that is central to studying gene regulation and determining protein:DNA interactions is the electrophoretic mobility shift assay (EMSA).
The EMSA technique is based on the observation that protein:DNA complexes migrate more slowly than free DNA molecules when subjected to non-denaturing polyacrylamide or agarose gel electrophoresis. Because the rate of DNA migration is shifted or retarded upon protein binding, the assay is also referred to as a gel shift or gel retardation assay. Until conception of the EMSA protein:DNA interactions were studied primarily by nitrocellulose filter-binding assays.
All that is needed to perform the assay is purified DNA target that has been end-labeled with biotin, the protein extract to be tested, nylon membrane and basic electrophoresis equipment. DNA targets can be synthesized with 5' or 3' biotin labels or they can be labeled after synthesis using the Thermo Scientific Biotin 3' End DNA Labeling Kit (Product No. 89818). Nuclear, cytosolic or whole cell protein extracts can be obtained by a variety of methods, including the Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Reagent Kit (Product No. 78833).
More Product Data
• Transfer EMSA gels using the Pierce G2 Fast Blotter
Related Products
LightShift™ Chemiluminescent EMSA Kit
LightShift™ Poly (dI-dC)
Features of theLightShift EMSA Optimization and Control Kit:
• 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
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.
The interaction of proteins with DNA is central to the control of many cellular processes including DNA replication, recombination and repair, transcription, and viral assembly. One technique that is central to studying gene regulation and determining protein:DNA interactions is the electrophoretic mobility shift assay (EMSA).
The EMSA technique is based on the observation that protein:DNA complexes migrate more slowly than free DNA molecules when subjected to non-denaturing polyacrylamide or agarose gel electrophoresis. Because the rate of DNA migration is shifted or retarded upon protein binding, the assay is also referred to as a gel shift or gel retardation assay. Until conception of the EMSA protein:DNA interactions were studied primarily by nitrocellulose filter-binding assays.
All that is needed to perform the assay is purified DNA target that has been end-labeled with biotin, the protein extract to be tested, nylon membrane and basic electrophoresis equipment. DNA targets can be synthesized with 5' or 3' biotin labels or they can be labeled after synthesis using the Thermo Scientific Biotin 3' End DNA Labeling Kit (Product No. 89818). Nuclear, cytosolic or whole cell protein extracts can be obtained by a variety of methods, including the Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Reagent Kit (Product No. 78833).
More Product Data
• Transfer EMSA gels using the Pierce G2 Fast Blotter
Related Products
LightShift™ Chemiluminescent EMSA Kit
LightShift™ Poly (dI-dC)
For Research Use Only. Not for use in diagnostic procedures.
Specifications
AssayEMSA Assay
FormatKit
Includes10X 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), 1M KCl (1 mL), 100mM MgCl2 (500 μL), 200mM EDTA pH 8.0 (500 μL), 5X Loading Buffer (1 mL)
For Use With (Equipment)myECL™ Imager, X-Ray Film
Product LineLightShift™
Product TypeLightShift EMSA Optimization and Control Kit
Quantity100 Reactions
Target SpecificityNot Target-Specific
TechniqueGel Shift
Unit SizeEach
Contents & Storage
Sufficient For: 100 binding reactions
• 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
Store at -20°C.
• 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
Store at -20°C.