Detects rat GFR alpha ‑1/GDNF R alpha ‑1 in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant human GFR alpha ‑1 is observed and less than 1% cross-reactivity with recombinant mouse GFR alpha ‑2 is observed.
Polyclonal Goat IgG
Mouse myeloma cell line NS0-derived recombinant rat GFR alpha ‑1/GDNF R alpha ‑1 Asp25-Leu445 Accession # Q62997
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Blockade of Receptor-ligand Interaction
In a functional ELISA, 4-10 µg/mL of this antibody will block 50% of the binding of 4 ng/mL of Recombinant Human GDNF (Catalog # 212-GD) to immobilized Recombinant Rat GFR alpha -1 Fc Chimera (Catalog # 560-GR) coated at 0.5 µg/mL (100 µL/well).
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
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 GFR alpha ‑1/GDNF R alpha ‑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 GFR alpha ‑1/GDNF R alpha ‑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. GFR alpha ‑1/GDNF R alpha ‑1 was detected in perfusion fixed frozen sections of rat spinal cord using Goat Anti-Rat GFR alpha ‑1/GDNF R alpha ‑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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: GFR alpha-1/GDNF R alpha-1
Glial cell line-derived growth factor (GDNF), neurturin (NTN) and persephin, distant members of the TGF-beta superfamily, are neurotrophic factors for a variety of neuronal populations in the central and peripheral nervous systems. The bioactivities of GDNF and NTN are mediated through a receptor complex composed of the non ligand-binding signaling subunit (c-Ret receptor tyrosine kinase) and either of two ligand binding subunits (GDNF receptor alpha - (GFR alpha -1) or GFR alpha -2). GFR alpha -1 and ‑2 are members of a family of at least four cysteine-rich glycosyl-phosphatidylinositol (GPI)-linked cell surface proteins that share conserved placements of many of their cysteine residues. Binding of GDNF to membrane-associated GFR alpha -1 or GFR alpha -2 initiates the association with and activation of the Ret tyrosine kinase. Soluble GFR alpha s released enzymatically from the cell surface-associated protein with phosphatidylinositol phospholipase C, as well as recombinantly produced soluble GFR alpha -1, can also bind with high-affinity to GDNF and trigger the activation of Ret tyrosine kinase. Rat GFR alpha -1 cDNA encodes a 468 amino acid (aa) residue protein with an N‑terminal 24 aa residue hydrophobic signal peptide. Like other GPI-linked proteins, rat GFR alpha -1 has a C-terminal hydrophobic region which is preceded by a three aa residue (ASS) GPI-binding site. Human GFR alpha -1 shares 93% amino acid identity with rat GFR alpha -1. The expression of the various GFR alpha s are differentially regulated in the central and peripheral nervous system, suggesting complementary roles for the GFR alpha s in mediating the activities of the GDNF family of neurotrophic factors.
Thompson, J. et al. (1998) Mol. Cell Neurosci. 11:117.
Trupp, M. et al. (1998) Mol. Cell Neurosci. 11:47.
Baloh, R.H. et al. (1998) Proc. Natl. Acad. Sci. USA 95:5801.
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Immunocytochemistry/Immunofluorescence: Rat GFR alpha ‑1/GDNF R alpha ‑1 Antibody [AF560]
Other Experimental Details
Other Experimental Details
Frozen sections from paraformaldehyde fixed postnatal day 1 mouse testes were cut at 5 microns and affixed to sides. Sections were permealbilized with 0.1% Triton X-100 in PBS, then blocked with 3% BSA in 0.1% Triton X-100 in PBS for 30 min. Sections were then incubated for 1 hour at RT with Goat Anti-Rat GFR alpha 1/GDNF R alpha 1 Antibody (R&D Systems AF560) at 5 µg/ml. After washing 3X with 0.1% Triton X-100 in PBS, sections were incubated with Donkey anti-Goat (H+L) Secondary Antibody, Alexa Fluor 555 conjugate (red; ThermoFisher Scientific A21432) for 1 hour at RT. Phalloidin was used a counterstain (blue) to label F-actin.