Living colors gfp monoclonal antibody

Antibodies used were: rabbit monoclonal anti-Src (#32G6, Cell Signaling Technologies, Danvers, MA) diluted 1:5000; rabbit polyclonal anti-Pyk2 (#3292, Cell Signaling Technologies, Danvers, MA) diluted 1:2000; Rabbit polyclonal anti-MBD2 (#ab224827, Abcam, Cambridge, MA diluted 1:1000; 3 additional MBD2 antibodies were used in (unsuccessful) efforts to detect FP-MBD2 including: rabbit anti-MBD2 (#NB100-81657, Novus Biologicals, Centennial, CO); mouse monoclonal anti-MBD2 (#N2C3, GeneTex, Irvine, CA); and rabbit polyclonal anti-MBD2 (#ab38646, Abcam, Cambridge, MA). GFP was detected using mouse monoclonal anti-GFP (Living Colors GFP monoclonal antibody, Clontech, Mountain View, CA) diluted 1:10,000; HRP-goat-anti-mouse IgG antibody or HRP rabbit-anti-mouse IgG (Jackson ImmunoResearch, West Grove, PA, USA) diluted 1:10,000. Protein A-sepharose was used to precipitate immune complexes in immunoprecipitation experiments.

Antibodies against ADAM12 were as previously described [23 (link),36 (link),37 (link)]. The rabbit polyclonal antibodies N-19 (sc-9752) and H-200 (sc-13976) against BSG, and goat polyclonal antibody L-19 against PLAP (sc-9757) were from Santa Cruz Biotechnology (Dallas, TX, USA). Other antibodies used were rabbit polyclonal antibody PA5-29787 against BSG (Thermo Fischer Scientific, Waltham, MA, USA), mouse monoclonal antibodies against actin (MAB1501, Millipore Chemicon, Burlington, MA, USA) and FITC-conjugated anti-CD147 (555962, BD Biosciences, San Jose, CA, USA). Living Colors^® GFP Monoclonal Antibody from Clontech was used for co-immunoprecipitation studies. Secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-mouse, goat anti-rabbit, and rabbit anti-goat immunoglobulins from DAKO A/S (Glostrup, Denmark). Alexa Fluor 488-conjugated rabbit anti-goat IgG and goat anti-mouse IgG, and Alexa Fluor 546-conjugated goat anti-mouse IgG F(ab)₂ fragment and goat anti-rabbit IgG F(ab)₂ fragment were from Invitrogen (Taastrup, Denmark). The metalloprotease inhibitors Batimastat, GM6001, and all other chemicals were from Merck KGaA (Darmstadt, Germany).