Goat antibody to Calretinin

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Publication protocol

Free-floating sections were incubated in goat anti-CR (1:1000; SWANT, Bellinoza, Switzerland; Schiffmann et al., 1999), rabbit anti-Pax2 (1:1000; Invitrogen, Paisley, United Kingdom; Dressler and Douglass, 1992), and mouse anti-HCN4 (1:500; UC Davis/NIH NeuroMab Facility, Davis, CA, United States; Giesbrecht et al., 2010; Khurana et al., 2012). Hyperpolarization-activated cation-4 labeling was visualized using a tetramethylrhodamine kit (PerkinElmer Life Sciences, Boston, MA, United States) for tyramide signal amplification as described previously (Hughes et al., 2012), whereas immunolabeling for CR and Pax2 was visualized using species-specific secondary antibodies conjugated to Alexa 647 and Pacific Blue, respectively. All primary and secondary antibody cocktails were made up in 0.3 M phosphate-buffered saline with 0.3% Triton X-100. Sections were incubated in primary antibodies for 72 h and in secondary antibodies for 12–18 h at 4°C.

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