Goat anti rabbit dylight 488 secondary antibody

Brains from Kiss1^Cre/+ and Kiss1^Cre/Cre mice (n = 5/group) injected with AAV5-hSyn-DIO-hM3D(Gq)-mCherry were assessed for mCherry reporter expression and GnRH to confirm anatomical integrity of Kiss1 neuron and GnRH fiber interaction in the area. A potential explanation for the lack of LH responses in DREADD-injected Kiss1 KO mice could be developmental alterations in the projections from KNDy neurons to GnRH neurons (Clarkson and Herbison, 2006 (link)). Furthermore, that could explain the lack of LH stimulation after chemogenetic activation of Kiss1 neurons in Kiss1 KO mice. The anatomical relationship between the two proteins appeared to be similar in both genotypes (Kiss1^Cre/+ and Kiss1 KO), suggesting that kisspeptin is not needed developmentally for the formation of fibers in Kiss1 neurons. (Figure 2—figure supplement 1). One series of free-floating sections containing ARC and amygdala from each animal was incubated overnight in blocking solution containing rat anti-mCherry primary antiserum, Alexa Fluor 594 conjugate (1:500; Thermo Fisher, Cat. No. M11240). The next morning sections were extensively washed in PBS and then incubated overnight in rabbit anti-GnRH (1:1,000; Abcam, Cat. No. ab5617) followed by goat-anti-rabbit DyLight 488 secondary antibody (1:200; Thermo Scientific, Cat. No. 35552).

Brains from Kiss1^Cre/+ and Kiss1^Cre/Cre mice (n = 5/group) injected with AAV5-hSyn-DIO-hM3D(Gq)-mCherry were assessed for mCherry reporter expression to confirm bilateral infection of ARC or MePD GFP expressing Kiss1 neurons with the AVV carrying Cre-dependent hM3D receptor. One series of free-floating sections containing ARC and amygdala from each animal was incubated overnight in blocking solution containing rat anti-mCherry primary antiserum, Alexa Fluor 594 conjugate (1:500; Thermo Fisher, Cat. No. M11240). The next morning sections were extensively washed in PBS and then incubated overnight in rabbit anti-GFP tag antibody (1:5,000; Thermo Fisher, Cat. No. A-6455) followed by goat-anti-rabbit DyLight 488 secondary antibody (1:200; Thermo Scientific, Cat. No. 35552).