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2 protocols using cdc37

1

Inhibition of Hsp90 and Proteasome in Cancer Cells

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Y‐632 was synthesized at the Shanghai Institute of Materia Medica, Chinese Academy of Sciences (Shanghai, China). Imatinib mesylate was obtained from Selleck Chemicals (Houston, TX, USA). Geldanamycin was obtained from Sangon Biotech (Shanghai, China). MG132, proteasome inhibitor I (PS341), chloroquine, MDL28170, monobromobimane, SRB, MTT, and antibodies against β‐tubulin and insulin‐like growth factor 1 receptor were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Antibodies against human epidermal growth factor receptor‐2, GAPDH, EGFR, c‐Met, Akt, phospho‐Akt (Ser473), Cdk4, Cdk6, p85, poly‐(ADP‐ribose) polymerase, caspase 8, caspase 9, Hop, Cdc37, Erk1/2, and phospho‐Erk1/2 (Thr202/Tyr204) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against β‐actin, Raf‐1, p23, and caspase 3 were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against Hsp90 and Bcr‐Abl were from Abcam (Cambridge, UK). Anti‐Hsp70 antibody was from Enzo Life Sciences (Farmingdale, NY, USA). Antiphosphotyrosine antibody (clone 4G10) was obtained from Millipore (Billerica, MA, USA).
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2

Comprehensive Antibody Panel for SGK1, AKT, Chk1, and NDRG1 Signaling

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Anti-SGK1 (#12103), AKT (#4691), Phospho-AKT-Ser473 (#9271), Phospho-AKT-Thr308 (#13038), Chk1(#2360), Phospho-Chk1-Ser345 (#2348), Phospho-Histone H2A.X-Ser139 (#2577), NDRG1(#9395), Phospho-NDRG1-Thr346 (#3217), Phospho-NEDD4L-Ser448 (#8063), Hsp90 (#4877), CDC37(#4793), HA Tag (#3724), and FLAG Tag (#14793) were from Cell Signaling Technology. Phospho-SGK1-S422 (#44-1264G), FAM122A (#MA5-24510), V5-HRP (#R961-25) were from Invitrogen; NEDD4-2 (#ab131167) from Abcam; PP5 antibody (#sc-271816) from Santa Cruz Biotechnology; FAM122A (#NBP2-31646) from Novus Biologicals; GAPDH-HRP (#BE0034-100) from Easybio; actin-HRP (#AM8502b) from ABGENT; and secondary antibodies, anti-rabbit IgG HRP (#A16110), and anti-mouse IgG HRP (#A16072) from Invitrogen.
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