Rat GFR alpha-1/GDNF R alpha-1 Antibody

Detection of Rat GFR alpha‑1/GDNF R alpha‑1 by Western Blot.

Western blot shows lysates of rat brain tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Rat GFRa-1/GDNF Ra-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF560) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for GFRa-1/GDNF Ra-1 at approximately 52 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.

GFR alpha‑1/GDNF R alpha‑1 in Rat Spinal Cord.

GFRa-1/GDNF Ra-1 was detected in perfusion fixed frozen sections of rat spinal cord using Goat Anti-Rat GFRa-1/GDNF Ra-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF560) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to spinal cord dorsal horn. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse GFR alpha-1/GDNF R alpha-1 by Immunocytochemistry/ Immunofluorescence

Ectopic RAR gamma expression by GFR alpha1+ spermatogonia. (A) The CAG-CAT-3xFLAG-Rarg transgene. When CAT between the loxP sites is deleted by TM-activated Cre, FLAG-tagged RAR gamma is constitutively expressed under the control of the CAG promoter. (B) Experimental design of the fate analysis of GFR alpha1+ cells with enforced FLAG-RAR gamma expression upon VA readministration in VAD mice, as shown in C-F. Gfra1-CreERT2; CAG-CAT-3xFLAG-Rarg transgenic mice were maintained in VAD and VA was administered 2 days after TM injection, as indicated. Testes were then processed for IF. (C,D) IF images of whole-mount seminiferous tubules of the mice described above, 2 days after VA injection, stained for FLAG-RAR gamma (green) and KIT (magenta) (C), and cell number relative to the number of initial induced cells (D). Data for GFP-labeled NGN3+ and GFR alpha1+ cells are reproduced from Fig. 2C and Fig. 3C, respectively, for comparison. The mean±s.e.m. value of three testes is shown. *P<0.003 (t-test), compared with the values of FLAG-RAR gamma+ GFR alpha1+ cells at day 2. (E,F) Representative confocal images of the same field of whole-mounts of seminiferous tubules of mice treated as described above, at 2 days after VA injection; staining was performed for GFR alpha1, KIT and FLAG (E). Open arrowheads, white arrowheads and small arrows indicate FLAG+ cells that are GFR alpha1+/KIT+, GFR alpha1+/KIT− and GFR alpha1−/KIT+, respectively. (F) Quantitation of GFP+ and FLAG-RAR gamma+ cells showing different patterns of GFR alpha1 and KIT expression in Gfra1-CreERT2; CAG-CAT-EGFP and Gfra1-CreERT2; CAG-CAT-3xFLAG-Rarg mice, respectively. Cell numbers are shown above each bar. (G) Experimental design of the fate analysis of GFR alpha1+ cells with enforced FLAG-RAR gamma expression under normal conditions, as shown in H-J. Gfra1-CreERT2; CAG-CAT-3xFLAG-Rarg transgenic mice were pulsed with TM at 13-17 weeks of age, and after 2 and 10 days their testes were processed for IF. (H) IF images of whole-mount seminiferous tubules 2 and 10 days after TM injection, stained for FLAG-RAR gamma and GFR alpha1. (I,J) Numbers of GFR alpha1+ Aundiff (magenta), GFR alpha1− Aundiff (green), KIT+ (blue) spermatogonia and total cells (black) in either GFP-labeled (I) or FLAG-RAR gamma-expressing (J) cells of Gfra1-CreERT2; CAG-CAT-EGFP and Gfra1-CreERT2; CAG-CAT-3xFLAG-Rarg mice, respectively, following the schedule shown in G. The mean±s.e.m. of four (I) and three (J) testes are shown. *P<0.05 (t-test), compared with the values on day 2. Scale bars: 50 μm. Image collected and cropped by CiteAb from the following open publication (https://journals.biologists.com/dev/article/doi/10.1242/dev.118695/258623/Hierarchical-differentiation-competence-in), licensed under a CC-BY license. Not internally tested by R&D Systems.