Anti pdk1 antibody

Cellular lysates were prepared by suspending 1 × 10⁶ cells in 100 µl of RIPA lysis buffer (1 × PBS, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS) that was supplemented with 10 mM β-glycerophosphate, 1 mM sodium orthovanadate, 10 mM NaF, and 1 mM phenylmethylsulfonyl fluoride. The cells were extracted on ice for 30 min. The proteins were electro-transferred to nitrocellulose membranes (Millipore Corp., Bedford, MA), and specific proteins were detected using chemiluminescence. Anti-PDK1 antibody (cat#: 10026-1-AP), anti-Akt antibody (cat#: 10176-2-AP), anti-p-Akt (S473) antibody (cat#: 66444-1-Ig), anti-SOD3 antibody (cat#: 14316-1-AP), anti- GAPDH antibody (cat#: 60004-1-Ig), anti-PRDX1 antibody (cat#: 15816-1-AP) were purchased from Proteintech Group, Inc, Rosemont, IL, USA. Anti-STAT3 antibody (cat#: 9139), anti-p-STAT3 (Y705) antibody (cat#: 9145), anti-cleaved caspase-3 antibody (cat#: 9664) were purchased from Cell Signaling Technology Inc., Danvers, MA, USA. All primary antibodies were diluted at 1:500.

Western blot was performed to measure protein expression levels of HIF-1α, PDK1, GLUT1, HK2, Ubiquitin, UFL1, and BRE1B. And following antibodies were used: anti- HIF-1α antibody (1:300; Proteintech, United States), anti-GLUT1 antibody (1:1,000; Proteintech, United States), anti-HK2 antibody (1:2000; Cell Proteintech, United States), anti-PDK1 antibody (1:2000; Proteintech, United States), anti-ubiquitin antibody (1:1,000; Proteintech, United States), anti-UFL1 antibody (1:1,000; Bethl Laboratories, United States) and anti- BRE1B antibody (1:1,000; Bethl Laboratories, United States).