Dylight488 goat anti rabbit secondary antibody

The staining of X‐gal and immunofluorescence was performed, as reported previously with slight modifications.⁸, ⁹, ¹⁰ Briefly, six sets of sequential coronary sections (40‐μm thickness) were collected. The brain sections were subjected to X‐gal staining solution (Sigma; BG‐3) overnight and then counterstained with 0.1% neutral red (Sigma; N4638) for 1 hour at 37°C. To analyze the proliferation and differentiation of cultured midbrain NPCs, each group of cells was incubated overnight with the rabbit anti‐doublecortin (DCX; 1:400; Cell Signaling Technology; 148025) on DIV 5, or rabbit anti‐β‐tubulin III (1:400; Cell Signaling Technology; 151155) on DIV 7, or guinea pig anti‐neuronal nuclear antigen (NeuN; 1:3000; Merck Millipore; ABN90) on DIV 15, or rabbit anti‐TH antibodies (1:400; Merck Millipore; AB152) on DIV16 alone or in combination with the mouse anti‐BrdU antibody (1:400; Novus; NB500‐169). The cells were then incubated with DyLight594 goat anti‐mouse or DyLight488 goat anti‐rabbit secondary antibody (1:400; Jackson ImmunoResearch) for 1.5 hours at 37°C. Finally, the cells were cover‐slipped with DAPI‐containing mounting medium (Sigma). Negative control was carried out by replacing with normal serum or vehicle solution.