Anti gapdh nb300 221

Recombinant human CCL5 (278‐RN‐050CF) was purchased from R&D Systems (Minneapolis, Minnesota, USA). The primary antibodies used in western blot and immunohistochemistry (IHC) assays included rabbit anti‐CCR5 (ab32048; Abcam, Cambridge, Massachusetts, USA), anti‐AR (sc‐7305; Santa Cruz Biotechnology, Dallas, Texas, USA) and anti‐GAPDH (NB300‐221; Novus Biologicals, Littleton, Colorado, USA). MAB‐678 antihuman CCL5 monoclonal neutralizing antibody was obtained from R&D Systems. Normal rabbit IgG (sc‐2027) used as a control in the CCL5 neutralizing antibody assay was obtained from Santa Cruz Biotechnology.

The cells were lysed by PRO‐PREP TM Protein Extraction Solution (iNtRON Biotechnology, Gyeonggi‐do, Korea) according to the manufacturer's instructions. The protein concentration was determined, and proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membranes. The membranes were blocked with BlockPro 1 min protein‐free blocking buffer and incubated with the following primary antibodies at 4°C overnight: anti‐NPRL2 (sc‐376986; Sigma‐Aldrich), anti‐NPRL3 (ab121346; Abcam, Cambridge, MA), anti‐DEPDC5 (GTX133570; GeneTex), anti‐Rag A (D8B5; Cell Signaling Technology), anti‐Rag C (D8H5; Cell Signaling Technology), anti‐phospho‐mTOR (Ser2448; Cell Signaling Technology), anti‐phospho–4E‐binding protein (4E‐BP1) (Thr37/46; 2855; Cell Signaling Technology), anti‐phospho‐S6K (Cell Signaling Technology), anti‐LC3 (NB‐100‐2220; Novus Biologicals), anti‐p62 (H0008878‐M01; Abnova), and anti‐GAPDH (NB‐300‐221; Novus Biologicals) antibody.