Mouse Ret Antibody

Ret in Mouse Spinal Cord.

Ret was detected in perfusion fixed frozen sections of mouse spinal cord using Goat Anti-Mouse Ret Antigen Affinity-purified Polyclonal Antibody (Catalog # AF482) 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 the ventral horn. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Porcine Ret by Western Blot

ERBB-family signaling molecules in rat testis cells. (a) Polypeptides in the EGF super-family signal by activating ERBB-family transmembrane receptor tyrosine kinases. ERBB1 is a receptor for ‘classical’ low molecular weight EGF-like peptides. ERBB2 is the primary transducer for ligand-bound ERBB1, ERBB3 and ERBB4. ERBB2’s extracellular domain does not bind known ligands. ERBB3 is a receptor for Neuregulin-1 (NRG1), NRG2 and Neuroglycan-C (CSPG5). Ligand bound ERBB3 displays poor kinase activity and signals most effectively as a heteromer with ERBB1, ERBB2 and/or ERBB4. ERBB4 is a receptor for NRG1, NRG2, NRG3 and NRG4 plus other EGF-like peptides*. (b) Western blotting analysis of ERBB-family proteins in fractions of testis cells from 23-day-old rats. Lysates of type A spermatogonia after proliferating for ~180 days/15 passages in culture (SgL), freshly isolated laminin-binding type A spermatogonia (Sg), laminin non-binding spermatogenic cells (Scy), tubular somatic cells (SC), interstitial somatic cells (IC), MCF7 human mammary gland cells (MCF) and COS7 monkey kidney cells (COS). Arrowheads: ERBBs 1–4 (~185 kDa), RET (~155 and 170 kDa) and TUBA1a (~55 kDa). (c) Relative abundance (qtPCR) of ERBB-family transcripts in testis cells isolated from 23-day-old rats (n=cells from three different rats; ±S.E.M.). Spermatogonia (Sg), Spermatocytes (Scy; differentiating spermatogonia/early spermatocytes), Tubular somatic cells (SC) and Interstitial somatic cells (IC) are cell types described in panel (b). (d) Testis cross-section from 26-day-old tgGCS-EGFP transgenic rats labeled with anti-ERBB2 (Red) overlaying EGFP fluorescence from germ cells (green). Note, cytoplasmic ERBB2 labeling in germ cells resembling differentiating spermatogonia (white arrows) and spermatocytes (yellow arrow). Scale, 40 μm. (e) Rat seminiferous tubule whole mount from 24-day-old wild-type rat labeled using antibodies to ERBB2 (Red) and ZBTB16 (Green). Scale, 20 μm. Note: nuclear ZBTB16 labeling is more robust in ERBB2-dim spermatogonia (cyan arrows), compared with ERBB2-bright spermatogenic cells (white arrows). (f) Rat seminiferous tubule whole mount from a 24-day-old wild-type rat labeled with antibodies to ERBB2 (Red) and phospho-Histone-3 (pH3, Green). Scale, 40 μm. Note: nuclear pH3 in large mitotic ERBB2+ syncytia. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/cddiscovery201518), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Ret by Immunohistochemistry

P2RX4 expression in Dorsal Root Ganglion neurons. (A) Representative cropped western blotting analysis of lumbar DRG extract indicates the presence of a 60 kDa band in P2RX4+/+ DRG that is absent in extracts from P2RX4−/− DRG. N = 3 independent experiments, n = 3 mice per genotype. (B) beta -galactosidase staining was used as a surrogate marker for P2RX4 in DRG sections. Only P2RX4−/− neurons are immuno-positive for beta gal compared to sections from P2RX4+/+ DRG. Scale bar 100 µm. (C) DRG neurons that express beta -gal innervate paw skin. Fluoro-Gold B, a retrograde marker of neurons was injected in paw-skin and L5/L6 DRG were removed for staining a week later. beta gal immunostaining reveals a population of neurons that are labeled for both beta gal and Fluoro-Gold B. Scale bar 50 µm. (D) In nociceptive neurons, beta gal is mainly expressed in c-ret-, TRPV1- and IB4-positive neurons, but scarcely in others populations (CGRP, TrkA or Substance P (SP) expressing cells). Representative images of DRG sections stained with beta gal and makers of nociceptive neurons. Scale bars 50 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29343707), licensed under a CC-BY license. Not internally tested by R&D Systems.