Alexa fluor 488 labeled igg secondary antibody

After transcardial perfusion (4 % paraformaldehyde) of deeply anesthetized mice, brains were removed, cryo-protected in 30 % sucrose, frozen in isopentane and sectioned (30 µm) on a cryostat. Sections were then stored free-floating, at 4°C and protected from light. Fluorescence from GFP and TdTomato expression was detected without antibody staining. Enkephalin was detected by using a rabbit anti methionine-enkephalin (1:100; Immunostar #20065) antibody in conjunction with an Alexa Fluor 488 labeled IgG secondary antibody (1:200; Jackson ImmunoResearch). Tyrosine hydroxylase (TH) was detected by using a rabbit anti-TH (1:2,000; Millipore #AB152) antibody in conjunction with an Alexa Fluor 488 labeled IgG secondary antibody (1:200; Jackson ImmunoResearch).