Anti tkt

Collected cells were homogenized in lysis buffer (5% SDS, 10 mM EDTA, 50 mM NaCl, 10 mM Tris–HCl). Protein concentrations were determined using pierce BCA protein assay (Thermo Fisher, Rockford, IL, USA). Proteins were resolved using SDS-polyacrylamide gel electrophoresis and transferred onto nitrocellulose membrane. Membranes were blocked in 5% milk, incubated with primary antibody at a concentration of 1:1000, then incubated with secondary antibody at a concentration of 1:10,000 and read using ECL regent (Bio-Rad, Richmond, CA, USA). Antibodies anti-GCLM, anti-GSS, anti-GCLC, anti-GSR, anti-GPX1, anti-SOD1, anti-SOD2, anti-TKT, anti-G6PD, anti-CGL and anti-Actin were purchased from Abcam. Anti-NRF2 and anti-CBS antibody was purchased from Cell Signaling Technology.

Unless otherwise stated, all chemicals were from Sigma-Aldrich Co., Ltd. Anti-Nrf2 (H300; sc-13032), Anti-Ub (sc-8017), anti-MKP-1 (sc-1102) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antiserum against aldoketo reductases 1C1 and C2 (AKR1C) was kindly provided by Professor John Hayes (University of Dundee, Dundee, UK). Anti-IDH1 (Cat#: 3997) were from Cell Signaling Technology (Danvers, MA, USA). Anti-TKT (Cat#: ab88438), anti-ME1 (Cat#: ab87974), and anti-G6PD (Cat#: ab993) were from Abcam (Cambridge, United Kingdom). Actin antibody (Cat#: R1207) and IgG antibody (Cat#: HA1011) were purchased from Hua-an (Hangzhou, Zhejiang, China). Antibodies against NQO1, HO-1 or AKR1B10 were raised in this laboratory and described previously [37 (link), 38 (link)].