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Gel Shift Assay Systems

Detect Sequence-Specific DNA-Binding Proteins

  • Core System includes controls and consensus oligonucleotides for AP2 and SP1
  • Regular System adds consensus oligos (see Overview below) to Core System contents
  • Oligonucleotides can be 5´ end-labeled or used unlabeled in competition assays

Choose a Gel Shift Assay

Size

Catalog number selected: E3050

$ 385.00 Your price: Log In

Gel Shift Assay Systems
Core System/100 reactions
$ 385.00
Your price: Log In
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Rapid Detection of DNA-Binding Proteins

The gel shift or electrophoretic mobility shift assay provides a simple and rapid method for detecting DNA-binding proteins. This method is widely used to study sequence-specific DNA-binding proteins such as transcription factors. The assay is based on the observation that protein:DNA complexes migrate more slowly through a nondenaturing polyacrylamide gel than free DNA fragments or double-stranded oligonucleotides. The gel shift assay involves incubating a purified protein or a complex mixture of proteins (such as nuclear or cell extract preparations) with a 32P end-labeled DNA fragment containing the putative protein binding site. The reaction products are then analyzed on a nondenaturing polyacrylamide gel. The specificity of the DNA-binding protein for the putative binding site is established by competition experiments using unlabeled DNA fragments or oligonucleotides containing a binding site for the protein of interest or other unrelated DNA sequences.

The Core System (Cat.# E3050) includes HeLa Nuclear Extract and SP1 and AP2 Consensus Oligos that can be used as positive controls and serve as a reliable system for obtaining experience with gel shift assays. In addition, the Core System contains T4 Polynucleotide Kinase and Kinase 10X Buffer for labeling oligonucleotides as well as Gel Shift Binding 5X Buffer. Cat.# E3300 contains all of the above plus consensus oligos for AP1, OCT1, CREB, NF-κB, and TFIID.

Application
  • Assay for DNA-binding proteins that bind to transcription factor consensus sequences or DNA sequences of your choice.

For more information, see the Protocols & Applications Guide.

Reference
  1. Ausubel, F. et al. (1989) In: Current Protocols in Molecular Biology, Vol. 1, Green Publishing Associates and Wiley-Interscience, Unit 12.2.

Protocols

Specifications

You are viewing: 100 reactions Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

Kinase 10X Buffer

C131B 1 x 100μl View Product

AP2 Consensus Oligonucleotide

E321B 1 x 35pmol

SP1 Consensus Oligonucleotide

E323B 1 x 35pmol

HeLa Nuclear Extract

E352A 1 x 40μl View Product

Gel Shift Binding 5X Buffer

E358A 1 x 200μl View Product

T4 Polynucleotide Kinase

M410A 1 x 100u View Product

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

See Protocol for detailed storage recommendations.

 

Specifications

You are viewing: 100 reactions Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

Kinase 10X Buffer

C131B 1 x 100μl View Product

AP1 Consensus Oligonucleotide

E320B 1 x 35pmol

AP2 Consensus Oligonucleotide

E321B 1 x 35pmol

TFIID Consensus Oligonucleotide

E322B 1 x 35pmol

SP1 Consensus Oligonucleotide

E323B 1 x 35pmol

OCT1 Consensus Oligonucleotide

E324B 1 x 35pmol

CREB Consensus Oligonucleotide

E328B 1 x 35pmol

NF-κB Consensus Oligonucleotide

E329B 1 x 35pmol

HeLa Nuclear Extract

E352A 1 x 40μl View Product

Gel Shift Binding 5X Buffer

E358A 1 x 200μl View Product

T4 Polynucleotide Kinase

M410A 1 x 100u View Product

SDS

Choose language:

Certificate of Analysis

Search for Specific Certificate:

View more results
No results
Loading…

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

See Protocol for detailed storage recommendations.

 

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