ChIP Assay of MLL and Epigenetic Markers

ChIP assay was conducted as described previously^{50 (link)}, with SABiosciences Corporation’s ChampionChIP One-Day kit (Qiagen, Frederick, MD) following the manufacturer’s protocol. Chromatin from THP-1 cells were cross-linked, sonicated into an average size of ~500 bp, and then immunoprecipitated with antibodies against the N’-terminal of MLL (MLL-N), the C’-terminal of MLL (MLL-C), EZH2, SIN3A, H3K27Me3 or IgG (Abcam, Cambridge, MA). Purified DNA was amplified by real-time qPCR using primers targeting the promoter of ALOX5 as described before^{32 (link)}.

Free full text: Click here

Wang Y., Skibbe J.R., Hu C., Dong L., Ferchen K., Su R., Li C., Huang H., Weng H., Huang H., Qin X., Jin J., Chen J, & Jiang X. (2017). ALOX5 exhibits anti-tumor and drug-sensitizing effects in MLL-rearranged leukemia. Scientific Reports, 7, 1853.

Publication 2017

ChampionChIP One-Day kit MLL-C SIN3A H3K27Me3

Alox5 Antibodies Chip assay Chromatin Ezh2 Primers Thp 1 cells

Corresponding Organization :

Other organizations : First Affiliated Hospital Zhejiang University, University of Cincinnati, University of Chicago

Top 5 similar protocols

Variable analysis

independent variables

Antibodies against the N'-terminal of MLL (MLL-N)
Antibodies against the C'-terminal of MLL (MLL-C)
Antibodies against EZH2
Antibodies against SIN3A
Antibodies against H3K27Me3

dependent variables

Binding of MLL-N, MLL-C, EZH2, SIN3A, H3K27Me3, and IgG to the promoter of ALOX5

control variables

THP-1 cells
Cross-linking of chromatin
Sonication of chromatin into ~500 bp fragments
Primers targeting the promoter of ALOX5

positive controls

None specified

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!