Alexa fluor 488 conjugated anti mouse igg h l

Mouse monoclonal antibody directed against human MET (#3148) and rabbit polyclonal antibody directed against the phosphorylated tyrosine (Y 1234/1235) of MET (#3126) were purchased from Cell Signaling Technology (Danvers, MA, USA). Mouse monoclonal antibody against the kinase domain of MET (3D4) was purchased from Invitrogen. Mouse monoclonal antibody directed against the N-terminal domain of MET (DL-21) was kindly provided by Dr Sylvia Giordano (University of Torino Medical School, Italy) [50 (link)]. Mouse monoclonal antibody against lamin B2 (33–2100) was purchased from Zymed. Mouse monoclonal antibody against GAPDH (6C5-32233), rabbit polyclonal antibody against the C-terminal domain of human PARP-1 (H250), and antibody against ERK2 (C14) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Green-fluorescent Alexa fluor 488 conjugated anti-mouse IgG (H+L) and phalloidin– Alexa Fluor 546 were purchased from Invitrogen. Rabbit polyclonal antibodies against the Flag epitopes were purchased from Sigma (St Louis, MO, USA).

Normal human skin sections were blocked with 1% BSA/PBS for 30 min and incubated with antibodies at RT for 1 hour. After washing with PBS, sections were incubated with Alexa Fluor 488-conjugated anti-mouse IgG (H + L) (Invitrogen) diluted 1:1000 with 1% BSA/PBS. The sections were observed with BX51 fluorescence microscope (Olympus, Tokyo, Japan).