Myoparr-binding proteins were collected with the RiboTrap Kit (Medical & Biological Laboratories (MBL), Aichi, Japan) using BrU labeled RNAs, as described previously [8 (link)]. BrU-labeled Myoparr and EGFP RNA were prepared using the Riboprobe System (Promega, Madison, WI, USA). Twenty-four hours after the induction of differentiation, nuclear extract was prepared from differentiating C2C12 myoblasts. The Myoparr or EGFP RNA (50 pmol) were mixed with the nuclear extract from differentiating C2C12 myoblasts for 2 h at 4 °C. The RNA–protein complexes were collected by Protein G Plus Agarose (Thermo Fisher Scientific, Waltham, MA, USA) conjugated to an anti-BrdU antibody, and proteins were eluted by adding BrdU. Purified proteins were detected by western blotting using a specific antibody, as described above.
Ribotrap kit
Manufactured by Medical & Biological Laboratories
Sourced in Japan
The RiboTrap Kit is a laboratory tool designed for the rapid and efficient isolation of ribonucleic acid (RNA) from various biological samples. It utilizes a selective binding matrix to capture and purify RNA molecules, enabling researchers to obtain high-quality RNA for subsequent analysis and applications.
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Lab products found in correlation
2 protocols using ribotrap kit
1
Myoparr-binding Protein Identification
2
Affinity Purification of Myoparr Interactors
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Myoparr-binding proteins were collected with the RiboTrap Kit (Medical & Biological Laboratories (MBL), Aichi, Japan) using BrU labeled RNAs, as described previously (Hitachi et al., 2019) . BrU-labeled Myoparr and EGFP RNA were prepared using the Riboprobe System (Promega, Madison, WI, USA). Twenty-four hours after the induction of differentiation, nuclear extract was prepared from differentiating C2C12 myoblasts. The Myoparr or EGFP RNA (50 pmol) were mixed with the nuclear extract from differentiating C2C12 myoblasts for 2 h at 4°C. The RNA-protein complexes were collected by Protein G Plus Agarose (Thermo Fisher Scientific) conjugated to an anti-BrdU antibody, and proteins were eluted by adding BrdU. Purified proteins were detected by Western blotting using a specific antibody, as described above.
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