Express one step sybr greener universal kit

F-12 Ham (HF-12) of Dulbecco’s modified Eagle’s medium, bovine serum albumin (BSA), trypsin-EDTA, Igepal CA-930, PGN from S. aureus 77140, Wortmannin (Wort), Akt-i IV, LY294002 (LY), SB216763 (SB), NaCl, LiCl, puromycin, and Bradford reagents were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Fetal calf serum (FCS) was acquired from Equitech-Bio, Inc. (Kerrville, TX, USA). A cocktail of sodium penicillin G, streptomycin sulfate, and amphotericin B was purchased from Gibco-BRL (Gaithesburg, MD, USA). Akt Inhibitor II, D-3-Deoxy-2-O-methyl-myo-inositol 1-[(R)-2-methoxy-3-(octadecyloxy) propyl hydrogen phosphate (SH-5) was acquired from Calbiochem (Darmstadt, Germany). Halt Phosphatase inhibitor cocktail was purchased from Thermo Fisher Scientific (Waltham, Massachusetts, USA). Protease inhibitor cocktail was acquired from GE Healthcare Bio-sciences (Little Chalfont, UK). Trizol reagent and EXPRESS One-Step SYBR GreenER Universal Kit were purchased from Invitrogen (Carlsbad, CA, USA). Bovine Interleukin 12 (IL-12/p40) TSZ ELISA kit was purchased from Biotang (Massachusetts, USA). All other reagents were acquired from Sigma-Aldrich.

DMEM/F12 (F12), Trypsin-EDTA, Igepal CA-930, Akt-IV, and SB216763 were purchased from Sigma–Aldrich, Inc. (St. Louis, MO, USA). Fetal calf serum (FCS) was acquired from Equitech-Bio, Inc. (Kerrville, TX, USA). A cocktail of sodium penicillin G, streptomycin sulfate, and amphotericin B was purchased from Gibco-BRL (Gaithersburg, MD, USA). Halt-TM Phosphatase inhibitor cocktail was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Protease inhibitor cocktail was acquired from GE Healthcare Biosciences (Little Chalfont, UK). Trizol reagent and EXPRESS One-Step SYBR GreenER Universal Kit were purchased from Invitrogen (Carlsbad, CA, USA). Bovine IL-10 and IL-8 TSZ ELISA kit were purchased from Biotang (Waltham, MA, USA). Plasmid pLKO.1-GSK3β-#1 was a gift from Alex Toker (Addgene plasmid #32497) and PLKO.1 plasmid was a gift from Bob Weinberg (Addgene plasmid # 8453). pCF CREB M1 was a gift from Marc Montminy (Addgene plasmid # 22969). The generation of pCF empty vector was done by the enzyme digestion of pCF CREB M1 with SacI restriction enzyme, and then the purification and ligation of the 5-kb fragment were generated.