Anti cxcl12 antibody

Total proteins were extracted from MTC tissues or cells by RIPA buffer (Abcam, MA, USA) supplemented with protease inhibitor cocktail. Protein concentration was quantified by BCA assay using the standard kit (Thermo Fisher Scientific, USA). Equal amounts of protein samples were loaded and separated through SDS-PAGE. Proteins in the gels were then transferred to Nitrocellulose membranes (Millipore, MA, USA). Following that, 5 % skimmed milk was used to block the membranes for half an hour at room temperature first and then primary antibodies were added to incubate at 4˚C overnight. The membranes were then washed with TBST 3 times and then incubated with corresponding secondary antibodies (KPL, USA) at room temperature for 2 h. The membranes were washed again before visualization by the ECL kit. Primary antibodies used for the study were listed as follows: anti-Snail antibody (1: 1000, Abcam, USA); anti-N-cadherin antibody (1: 1000, Abcam, USA); anti-E-cadherin antibody (1: 1000, Abcam, USA); anti-Vimentin (1: 1000, Abcam, USA); anti-CXCL12 antibody (1: 1000, Abcam, USA); anti-GAPDH (1:2000, Abcam, USA).

Eight-wk-old C57/B6 mice were sacrificed with mandibles and femurs freshly dissected followed by 4%-formalin fixation. Tissue sections were incubated with anti-Cxcl12 antibody (1:400, Abcam, Cambridge, MA) overnight at 4 °C followed by secondary antibodies, Alexa Fluor® 647 Goat Anti-Rabbit IgG (H + L) Antibody (1:2000, Invitrogen).