Protocol detail

HMGA1 Interactome Characterization

Find Similar Protocols

Immunoprecipitations of HMGA1-FLAG fusions were performed from nuclear extracts of transiently transfected HEK293 cells using M2 anti-FLAG agarose (Sigma) as recommended by the manufacturer. Endogenous HMGA1 was immunopurified from nuclear extracts using a specific anti-HMGA1 antibody (Abcam). Immunopurifications with boiled anti-HMGA1 IgG were used as a control. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and blotting were performed as described previously (22 (link)). Antibodies used in western blot analyses were M2 anti-FLAG antibody (Sigma), anti-HMGA1 antibody (Abcam), anti-CTIP2 antibody (Abcam), anti-Cdk9 antibody (Pierce), anti-CycT1 antibody (Proteintech), anti-HEXIM1 antibody (Proteintech) and anti-LARP7 antibody (Proteintech) with the corresponding secondary horseradish peroxidase-coupled antibodies (Sigma) as recommended by the manufacturer.

Free full text: Click here

Eilebrecht S., Le Douce V., Riclet R., Targat B., Hallay H., Van Driessche B., Schwartz C., Robette G., Van Lint C., Rohr O, & Benecke A.G. (2014). HMGA1 recruits CTIP2-repressed P-TEFb to the HIV-1 and cellular target promoters. Nucleic Acids Research, 42(8), 4962-4971.

Publication 2014

M2 anti-FLAG agarose anti-HMGA1 antibody M2 anti-FLAG antibody

Agarose Anti antibody Anti igg Antibodies Cdk9 Hek293 cells Hmga1 Horseradish peroxidase Immunoprecipitations Sds page Western blot analyses

Top 5 similar protocols

Variable analysis

independent variables

Transient transfection of HEK293 cells
Immunopurification using M2 anti-FLAG agarose or anti-HMGA1 antibody

dependent variables

Presence/absence of HMGA1-FLAG fusions and endogenous HMGA1 in nuclear extracts
Presence/absence of CTIP2, Cdk9, CycT1, HEXIM1, and LARP7 in immunopurified complexes

control variables

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and blotting
Antibodies used in western blot analyses

positive controls

Immunopurification with M2 anti-FLAG agarose to detect HMGA1-FLAG fusions
Immunopurification with anti-HMGA1 antibody to detect endogenous HMGA1

negative controls

Immunopurification with boiled anti-HMGA1 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!