Analyzing AP-1 Transcription Factor Binding

EMSA were performed as described [17 (link)]. Briefly, nuclear extracts from cerebral cortex of E14.5 c-fos +/+ and c-fos −/− embryos were prepared as specified by Wang et al. [11 (link)] and stored at −70°C until use. Double stranded oligonucleotide containing the AP-1 consensus sequence TGAGTCA (5´-CGCTTGATGAGTCAGCCGGAA-3´) (Promega) was end-labeled with ³²P-ATP using T4 polynucleotide kinase. AP-1 binding reaction, AP-1 competition assay and electrophoresis in non-denaturing polyacrylamide gels were performed as stated by the manufacturer (Promega). For detection of the components of the AP-1 complex we used 1 uL of anti c-Fos (sc-52, Santa Cruz), anti Fra-2 (sc-604, Santa Cruz), anti c-Jun (sc-1694, Santa Cruz) and 0.5 uL of goat anti mouse (A11031, Molecular Probes).

Free full text: Click here

Velazquez F.N., Prucca C.G., Etienne O., D'Astolfo D.S., Silvestre D.C., Boussin F.D, & Caputto B.L. (2015). Brain development is impaired in c-fos −/− mice. Oncotarget, 6(19), 16883-16901.

Publication 2015

anti c-Fos (sc-52, anti c-Jun (sc-1694, A11031

Assay C fos Cerebral cortex Consensus sequence Embryos Emsa Fra 2 Goat Molecular probes Mouse Non denaturing polyacrylamide gels electrophoresis Oligonucleotide Polynucleotide kinase Promega

Corresponding Organization :

Other organizations : Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de Córdoba, Research Centre in Biological Chemistry of Córdoba, CEA Paris-Saclay - Etablissement de Fontenay-aux-roses, Université Paris Cité, Université Paris-Saclay, Institut de Radiobiologie Cellulaire et Moléculaire, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Inserm

Top 5 similar protocols

Variable analysis

independent variables

Genotype (c-fos +/+ and c-fos -/-)

dependent variables

AP-1 binding activity

control variables

Nuclear extracts from cerebral cortex of E14.5 embryos
Double-stranded oligonucleotide containing the AP-1 consensus sequence TGAGTCA
AP-1 binding reaction, AP-1 competition assay, and electrophoresis in non-denaturing polyacrylamide gels were performed as stated by the manufacturer (Promega)
Antibodies used for detection of AP-1 complex components (anti c-Fos, anti Fra-2, anti c-Jun, and goat anti mouse)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!