Muscle Transcription Factor Regulation Profiling

In vivo skeletal muscle ChIP was performed with sonicated nuclear extract prepared from formaldehyde-cross-linked gastrocnemius muscle according to [137 (link)]. For immunoprecipitation, we used anti-BMAL1 (ab93806, Abcam), anti-REV-ERBα (generous gift from Ron Evans), anti-RNA Polymerase II (#MMS-126R; clone 8WG1, Biolegends), anti-NCOR1 (#20018-1-AP, Protein Tech), anti-HDAC3 (ab7030, Abcam), or rabbit IgG (2027x, Santa Cruz). DNA was column purified and used for sequencing or real-time qPCR (enrichment expressed as fold-change relative to IgG; primer sequences used are listed in S4 Table).

Free full text: Click here

Dyar K.A., Hubert M.J., Mir A.A., Ciciliot S., Lutter D., Greulich F., Quagliarini F., Kleinert M., Fischer K., Eichmann T.O., Wright L.E., Peña Paz M.I., Casarin A., Pertegato V., Romanello V., Albiero M., Mazzucco S., Rizzuto R., Salviati L., Biolo G., Blaauw B., Schiaffino S, & Uhlenhaut N.H. (2018). Transcriptional programming of lipid and amino acid metabolism by the skeletal muscle circadian clock. PLoS Biology, 16(8), e2005886.

Publication 2018

ab93806 ab7030

Chip Clone Formaldehyde Gastrocnemius muscle Immunoprecipitation Ncor1 Primer Protein Rabbit Rna polymerase ii Skeletal muscle

Corresponding Organization : Ludwig-Maximilians-Universität München

Other organizations : University of Graz, University of Padua, Città della Speranza Foundation, University of Trieste

Top 5 similar protocols

Variable analysis

independent variables

Antibody used for immunoprecipitation (anti-BMAL1, anti-REV-ERBα, anti-RNA Polymerase II, anti-NCOR1, anti-HDAC3, or rabbit IgG)

dependent variables

DNA enrichment (expressed as fold-change relative to IgG)

control variables

Formaldehyde-cross-linked gastrocnemius muscle
Sonicated nuclear extract

controls

Positive control: Immunoprecipitation with antibodies against specific proteins (BMAL1, REV-ERBα, RNA Polymerase II, NCOR1, HDAC3)
Negative control: Immunoprecipitation with rabbit IgG

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!