mTOR Inhibitor XI, Torin1 - Calbiochem

Protocol tips

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Highly potent inhibitor for mTOR and DNA-PK while inhibits PI3-K only at much higher concentration	Culture the cells in DMEM medium containing 250nM Torin1

Upstream tips
Highly potent inhibitor for mTOR and DNA-PK while inhibits PI3-K only at much higher concentration
Protocol tips
Culture the cells in DMEM medium containing 250nM Torin1

Publication protocol

Proteins were separated on SDS‐PAGE electrophoresis gel or on NuPAGE Bis‐Tris gel (Invitrogen) and electroblotted onto nitrocellulose (Protran; Schleicher & Schuell). Blots were incubated with primary antibodies in 5% nonfat dry milk in TBS plus 0.1% Tween‐20 overnight at 4 °C. Detection was achieved using horseradish peroxidase‐conjugated secondary antibody (Bio‐Rad Laboratories) and visualized with ECL plus (GE Healthcare). The following primary antibodies were used in this study: anti‐p70S6K and anti‐phospho‐p70S6K (Thr389), anti‐4EBP1 and anti‐phospho‐4EBP1 (Thr37/46), anti LC3, anti‐ SNAIL, anti‐SLUG (Cell Signaling), polyconal anti‐p62, anti‐Beclin1 (Santa Cruz), anti‐Actin and anti β‐tubulin (Sigma).

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