Rat anti tra98 antibody

For immunofluorescence staining, tissues fixed with 4% paraformaldehyde in PBS were cryo-embedded in OCT compound (Sakura Finetechnical, Japan) and cut into 7-µm-thick sections. Nuclei were counterstained with Hoechst 33342 dye. Specimens were observed with a microscope (Olympus IX 73; Olympus, Tokyo, Japan). The following were used as primary antibodies: chicken anti-GFP antibody (1:1,000, Nakalai Tesque, Inc., Kyoto, Japan), rat anti-TRA98 antibody (1:200, Abcam, USA), and anti-GFRα1 antibody (1:200, R&D Systems, USA). The secondary antibodies used were mouse anti-chicken IgG and goat anti-rat IgG, conjugated with Alexa 488 or Alexa 555 (1:200; Life Technologies, USA), and donkey anti-goat IgG conjugated with Alexa 647 (1:200; Life Technologies, USA). For the detection of EdU, sections were treated with the Click-iTTM EdU Alexa FluorTM 555 Imaging Kit (Invitrogen: C10338) before exposure to the primary and secondary antibodies on immunohistochemical examination.

Tissues fixed with 4% paraformaldehyde in PBS were cryoembedded in OCT compound (Sakura FineTechnical) and cut into 7‐µm‐thick sections. Nuclei were counterstained with Hoechst 33 342 dye. Specimens were observed with a microscope (Olympus IX 73; Olympus). The following were used as primary antibodies: chicken anti‐GFP antibody (1:1000; Nacalai Tesque, Inc), rat anti‐TRA98 antibody (1:200; Abcam), and anti‐GFRα1 antibody (1:200; R&D Systems). The secondary antibodies used were mouse anti‐chicken IgG and goat anti‐rat IgG, conjugated with Alexa 488 or Alexa 555 (1:200; Life Technologies).