Anti tubulin sc 73242

The following primary antibodies were used: The mouse antiSIRT7 (sc-365344), rabbit anti-SIRT7 (sc135055), anti-p53 (FL) (sc-6243), anti-p53 (DO-1) (sc-126), anti-p21 (sc-271532), anti-actin (sc-8432) and anti-tubulin (sc-73242) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The anti-STRAP (18277-1-AP) was purchased from Proteintech (Wuhan, China), the anti-GAPDH (#5174S) was purchased from Cell Signaling Technology (Danvers, MA, US), the anti-FLAG (F3165) was purchased from Sigma–Aldrich (St. Louis, MO, US), the anti-HA (#26183) was purchased from Pierce (Rockford, Illinois, US), the anti-pan-acetyllysine (#9441L) was purchased from Cell Signaling Technology (Danvers, MA, US).pcDNA3.1-Flag/HA-STRAP/SIRT7 was obtained by subcloning STRAP/SIRT7 sequence into pcDNA3.1-Flag/HA vector. The mutant constructs were generated by PCR amplification of STRAP/SIRT7 with specific primers. STRAP full-length (FL), STRAP (ΔC1, ΔC3, ΔC4, ΔC5, ΔC6, WD3, WD4, WD6, WD7, CT) were subcloned into the pGEX4T-3 vector.
STRAP siRNA 1#: gggugcaacacugaauaag; siRNA 2#: uuacgcauauaugacuuga (Genepharma, Suzhou, China).

Cells were collected and lysed using RIPA buffer (150 mM NaCl, 1% IGEPAL® CA-630, 0.5% sodium deoxycholate, 0.1% SDS and 50 mM Tris-HCl pH 8) supplemented with protease and phosphatase inhibitors (Roche, Switzerland) and total protein concentration were measured by BCA assay. Whole-cell lysates (~20–30 μg of proteins) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Immunoblotting was performed using the following primary antibodies: anti-p38 (#8690), anti-phospho p38 (T180/Y182 - #4511), anti-p53 (#2524), anti-phospho pS6 (S235/236 - #4058), anti-Akt (#9272), anti-phospho-Akt (S473, #4060), anti-caspase-3 (clone 8G10; #9665), anti-HK2 (#2867), anti-HER2 (#2165), anti-mTOR (#2983), anti-phospho-mTOR (S2448—#5536), anti-PARP (#9542) from Cell Signaling Technology (Massachusetts, USA); anti-LC3 (L7543) from Sigma-Aldrich; anti-Actin (sc-8432), anti-GAPDH (sc-166545), anti-IκB-α (sc-371), anti-Lamp2 (sc-18822) and anti-tubulin (sc-73242) from Santa Cruz Biotechnology (Texas, USA); and anti-Hsc70 (10654–1-AP) from Proteintech (Illinois, USA).

Anti-HDAC6 (7612), mouse monoclonal anti-Myc-Tag (2276), rabbit polyclonal anti-Myc-Tag antibody (2278), anti-Cullin 3 (2759), anti-LC3-B (2775), EMT antibody sampler kit (9782), anti-Ac-α-Tubulin (5335) and anti-GST (2625) antibodies were purchased from Cell Signaling. Anti-TRIM24 (TIF1α, SC-271266), anti-Cullin 1 (SC-11384), anti-HA antibody (SC-805), anti-Tubulin (SC-73242) and anti-p27 (SC-527) antibodies were purchased from Santa Cruz. Anti-SPOP antibody (16750-1-AP) was purchased from Proteintech. Anti-GFP (8371-2) antibody was purchased from Clontech. Polyclonal anti-Flag antibody (F7425), monoclonal anti-Flag antibody (F-3165, clone M2), anti-vinculin antibody (V-4505), peroxidase-conjugated anti-mouse secondary antibody (A-4416), peroxidase-conjugated anti-rabbit secondary antibody (A-4914), anti-HA agarose beads (A-2095) and anti-Flag agarose beads (A-2220) were purchased from Sigma. All antibodies were used in 1:1000 dilutions in 5% non-fat milk for western blot.
MLN4924 was a kind gift from Dr. William Kaelin (Dana-Farber cancer institute). Bafilomycin A1 was kindly provided by Dr. Junying Yuan (Harvard Medical School). MG132 (BML-PI102-0005) was purchased from Enzo life science.