Antibodies of α tubulin

The chemical reagents of EVO, N-acetyl-l-cysteine (NAC), SP600125, U0126, BCIP, 3-(4,5,-dimethylthiazol)-2-yl-2,5-diphenyltetrazolium bromide (MTT), and NBT were obtained from Sigma Chemical (St. Louis, MO, USA). Antibodies of α-tubulin, poly(ADP ribose) polymerase (PARP), caspase-3, caspase-9, Bcl-2, and Bax were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies of total (t) and phosphorylated (p) MAPKs (tERK/pERK and tJNK/pJNK), and cyclinB1/cdc25c proteins were obtained from Cell Signaling Technology (Danvers, MA, USA). The colorigenic synthetic peptide substrates, Ac-DEVD-pNA (a caspase-3 substrate), Ac-YVAD-pNA (a caspase-9 substrate), and Ac-IETD-pNA (a caspase-8 substrate) were purchased from Calbiochem. Other chemicals not mentioned above were obtained from Sigma Chemical.

The chemical reagents of EVO, SP600125, BCIP, 3-(4,5,-dimethylthiazol)-2-yl-2,5-diphenyltetrazolium bromide (MTT), and NBT were obtained from Sigma Chemical (St. Louis, MO, USA). Antibodies of α-tubulin, poly (ADP ribose) polymerase (PARP), Casp-3, Casp-9, Bcl-2, and Bax were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies of total (t) and phosphorylated (p) MAPK (total extracellular signal-regulated kinase (ERK)/p-ERK and total c-Jun N-terminal kinase (JNK)/p-JNK) proteins were obtained from Cell Signaling Technology (Danvers, MA, USA). Other chemicals not mentioned above were obtained from Sigma Chemical. EVO-related chemicals were based on the coupling of 3,4-dihydro-β-carboline with substituted N-alkyl isatoic anhydride in pyridine. 3,4-Dihydro-β-carboline was prepared by reacting tryptamine with ethyl formate, followed by intramolecular ring closure in the presence of POCl₃. Their purities were >95% when analyzed by high-performance liquid chromatography (HPLC) [8 (link), 13 (link)].