Alpha tubulin polyclonal antibody

Plasmids encoding short hairpin RNAs (shRNA) against HOXB9 were synthesized by Genepharma Company (Shanghai, China). In a 6-well plate, cells were seeded at a density of 1×10⁵ cells/well. We followed the manufacturer’s instructions to transfect the cells with the shRNA plasmids and vectors, using Lipofectamine^™ 3000. The western blotting assay was conducted following the conventional experimental procedures. Briefly, the protein was separated by SDS-PAGE and then transferred to nitrocellulose membranes, which were incubated with the primary rabbit polyclonal antibody of HOXB9 protein (1:300, SANTA CRUZ) and Alpha Tubulin Polyclonal antibody (1:1000, Proteintech) at 4° C overnight.

Western blotting was performed following previous laboratory procedures [15 (link), 28 , 39 (link)]. The following primary antibodies were used for blotting: anti-parkin antibody (Abcam, catalog no. ab77924; Santa Cruz Biotechnology, catalog no. sc-32282), PINK1 antibody (Novus Biologicals, catalog no. BC100–494), caspase 3 polyclonal antibody (Proteintech, catalog no. 19677-1-AP), anti-optineurin antibody [EPR20654] (Abcam, catalog no. ab213556), phospho-TBK1/NAK (Ser172) (D52C2) XP rabbit monoclonal antibody (Cell Signaling Technology, catalog no. 5483), LC3 rabbit polyclonal antibody (Proteintech, catalog no. 14600-1-AP), TOMM40 rabbit polyclonal antibody (Proteintech, catalog no. 18409-1-AP), SOD2 rabbit polyclonal antibody (Proteintech, catalog no. 24127-1-AP), COXIV rabbit polyclonal antibody (Proteintech, catalog no. 11242-1-AP), GAPDH monoclonal antibody (Proteintech, catalog no. 60004-1-Ig), beta-actin monoclonal antibody (Proteintech, catalog no. 66009-1-Ig), and alpha-tubulin polyclonal antibody (Proteintech, catalog no. 11224-1-AP). Secondary antibodies included HP-goat anti-mouse (ZsBio, catalog no. ZB-2305Beijing, China) and HP-goat anti-rabbit (ZsBio, catalog no. ZB-2301). The blots were visualized using a 5200 Chemiluminescent Imaging System (Tanon Science and Technology).

BCA PROTEIN ASSAY KIT,MEMBRANE AND CYTOSOL PROTEIN EXTRACTION KIT,TRYPSIN–EDTA SOLUTION,BEYOCOLOR PRESTAINED COLOR PROTEIN MARKER,SDS-PAGE SAMPLE LOADING BUFFER,BEYOECL MOON were purchased from BEYOTIME.PHOSPHO-AMPKa (Thr172) RABBIT mAb and AMPKa ANTIBODY were acquired from CST.STREPTOZOCIN,PVDF MEMBRANCRS,PROTIEN G–AGAROSE, FOETAL BOVINE SERUM and COLLAGENASE were obtained from SIGMA.PERCOOL, SODIUM CITRATE BUFFER and PROTEINASE K were purchased from SOLARBO.GLUT3 ANTIBODY was procured from SCB.2-NBDG was bought from THERMOFISHER.AICAR was acquired from SELLECK.CN.HRP-CONJUGATED AFFNIPURE GOAT ANTI-MOUSE IgG(H + L),HRP-CONJUGATED AFFNIPURE GOAT ANTI-RABBIT IgG(H + L),FITC-CONJ UGATED AFFNIPURE GOAT ANTI-RABBIT IgG(H + L),R-PE-CONJUGATED GOAT ANTI-RABBIT IgG(H + L),CYTOKERATIN 7-SPECIFIC POLYCLONAL ANTIBODY, AMPK ALPHA MONOCLONAL ANTIBODY and ALPHA TUBULIN POLYCLONAL ANTIBODY were purchased from PROTEINTECH.ACRYL/BIS 30% SOLUTION, 1MTRIS- HClSOLUTION(pH6.8),4XTRIS-HCL/SDS(pH8.8),TEMED,TRIS–GLYCINE-SDSRUNNING BUFFER,EZ-BUFFERS C 10X WESTERN TANSFER BUFFER and EZ-BUFFERS H 10X TBST BUFFER were acquired from SANGON BIOTECH. PRIMSCRIP RT MASTER MIX and TB GREEN PREMIX EX TAQ were obtained from TAKARA.