Alkaline phosphatase alp activity kit

Sitagliptin was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The ERK1/2 inhibitor (PD98059) and FITC-conjugated anti-α-smooth muscle actin (α-SMA) monoclonal antibody were obtained from Sigma-Aldrich (St Louis, MO, USA). Mouse monoclonal core-binding factor α 1 (cbfa1) (ab54868) and mouse monoclonal α-SMA antibody (ab7817) were obtained from Abcam (Cambridge, MA, USA). Trypsin, fetal bovine serum (FBS) and Dulbecco's Modified Eagle's Medium (DMEM) were purchased from Gibco (of Thermo Fisher Scientific, Waltham, MA, USA). Rabbit polyclonal antibody against ERK1/2 and a mouse monoclonal antibody against p-ERK1/2 were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Secondary antibodies were purchased from Vector Labs (Burlingame, CA, USA). Western blot bands were detected using the ECL Advance Western blotting detection kit from GE Healthcare (Chalfont St Giles, UK). Cell counting kit-8 was purchased from Dojindo Molecular Technologies (Rockville, MD, USA). Transwell plates were purchased from Millipore (Bedford, MA, USA). Annexin V-FITC Kit was purchased from BD Biosciences (San Jose, CA, USA). The alkaline phosphatase (ALP) activity kit was from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Calcium Colorimetric Assay Kit was obtained from Biovision (Mountain View, CA, USA).

A7r5 VSMCs were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Dulbecco’s modified Eagle’s Medium, Nutrient Mixture F-12 (DMEM/F12) was obtained from Gibco (Grand Island, USA). CML was acquired from Polypeptide Laboratories (San Diego, USA). Streptozotocin (STZ) was obtained from Sigma–Aldrich Co. LLC (St. Louis, USA). A CML ELISA kit was obtained from Meixuan Biological Science and Technology Co., Ltd (Shanghai, China). A calcium assay kit and alkaline phosphatase (ALP) activity kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). A Von Kossa staining kit was purchased from Shunbai Biologicals Inc. (Shanghai, China). An Annexin V-FITC apoptosis detection kit was obtained from Sigma-Aldrich Co. LLC (St. Louis, USA). Glyceraldehyde-phosphate dehydrogenase (GAPDH) was acquired from Cell Signaling Technology, Inc. (Boston, USA). Antibodies against ALP, bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (Runx2) and all secondary antibodies were from Santa Cruz Biotechnology (Santa Cruz, USA).