Anti gapdh monoclonal antibody

Sphingosylphosphorylcholine (SPC) was purchased from Biomol (Plymouth Meeting, PA, USA). Tamoxifen was purchased from Funakoshi (Tokyo, Japan). Corn oil was purchased from Sigma Aldrich (St. Louis, MO, USA).
The following antibodies were used: mouse monoclonal anti-paxillin antibody (Cat#610620, BD Biosciences, San Jose, CA, USA), anti-paxillin polyclonal antibody (H85, Santa Cruz, Dallas, CA, USA), anti-GAPDH monoclonal antibody (Wako, Osaka, Japan), anti-phospho-MYPT1 (T850) with phosphorylation site corresponding to T853 in human MYPT1 (Milipore, Burlington, CA, USA), anti-MYPT1 antibody (H130, Santa Cruz, Dallas, CA, USA), anti-phospho-myosin light chain 2 (S19) monoclonal antibody (Cell Signaling, Danvers, MA, USA), anti-myosin light chain (20 kDa) monoclonal antibody (Sigma Aldrich, St. Louis, MO, USA), anti-α-smooth muscle actin antibody (Thermo Fisher, Waltham, MA, USA), anti-mouse Alexa Fluor 488 (Thermo Fisher, Waltham, MA, USA), and anti-rabbit Alexa Fluor 546 (Thermo Fisher,Waltham, MA, USA). Secondary HRP-labeled antibodies (anti-mouse and anti-rabbit) were purchased from Promega (Madison, WI, USA).

n-3 DPA (purity ≧ 98%) solution in 100% ethanol was obtained from Cayman Chemical (Ann Arbor, MI, USA). n-6 DPA (purity > 99%) was purchased from Nu-Chek Prep, Inc. (Elysian, MN, USA). DPA was dissolved in 100% ethanol to make a 30 mM stock solution. The stock solutions were stored at −20 °C, then diluted to final concentrations before use. Sphingosylphosphorylcholine (SPC) was purchased from Biomol (Plymouth Meeting, PA, USA). Fura-2/AM was purchased from Dojindo (Kumamoto, Japan). Bradykinin (BK) was purchased from Peptide institute, Inc. (Osaka, Japan). All other chemicals were purchased from Katayama Chemical (Osaka, Japan).
The following antibodies were used: anti-phospho-MYPT1 (Thr850) with phosphorylation site corresponding to Thr853 in human MYPT1 (Upstate), anti-GAPDH monoclonal antibody (Wako), anti-phospho-myosin light chain 2 (Ser19) monoclonal antibody (Cell Signaling), anti-myosin light chain (20 kDa) monoclonal antibody (Sigma Aldrich), anti-Rho-kinase-2 polyclonal antibody (H85, Santa Cruz), and anti-rabbit Alexa Fluor 488 (Thermo Fisher). Secondary HRP-labeled antibodies (anti-mouse and anti-rabbit) were purchased from Promega.

Rabbit polyclonal anti-Slc22a14 antibody was raised against a keyhole limpet hemocyanin-conjugated peptide corresponding to the C-terminal region of mouse Slc22a14 (⁶¹⁵PKMDLPVQSLKAQPP⁶²⁹). Peptide synthesis, animal immunisation, and collection of sera were performed by Sigma-Aldrich Japan. The antibody was purified by affinity chromatography using an antigen peptide-conjugated column. Anti-GAPDH monoclonal antibody, anti-human septin 4 (N) antibody, and anti-phosphorylated tyrosine antibody (4G10) were purchased from WAKO Pure Chemical Industries, Immuno-Biological Laboratories (IBL), and Merck Millipore, respectively. Horseradish peroxidase-conjugated and Alexa Fluor-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology and Life Technologies, respectively.