Anti-GDNF Antibody

Glial-derived neurotrophic factor, hGDNF, Astrocyte-derived trophic factor
    Cat #: ANT-014
    Alternative Name Glial-derived neurotrophic factor, hGDNF, Astrocyte-derived trophic factor
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide (C)DDNLVYHILRKHSAKR, corresponding to amino acid residues 192-207 of human GDNF (precursor) (Accession P39905). C-terminus of the mature GDNF.
    • Anti-GDNF Antibody
    Accession (Uniprot) Number P39905
    Gene ID 2668
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, dog, monkey, bovine - identical; rat 15/16 amino acid residues identical.
    RRID AB_2039876.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype Rabbit IgG.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 μl, 50 μl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Negative control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, if, ih, wb
    May also work in: ip*
    Western blot
    • Anti-GDNF Antibody
      Western blot analysis of Recombinant human GDNF protein:
      Lanes 1, 4: 50 ng Recombinant human GDNF protein (#G-240).
      Lanes 2, 5: 10 ng Recombinant human GDNF protein.
      Lanes 3, 4: 1 ng Recombinant human GDNF protein.
      Lanes 1-3: Anti-GDNF Antibody (#ANT-014), (1:1000).
      Lanes 4-5: Anti-GDNF Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    • Anti-GDNF Antibody
      Expression of GDNF in rat dorsal root ganglion (DRG).
      Immunohistochemical staining of rat DRG longitudinal frozen section using Anti-GDNF Antibody (#ANT-014), (1:50), followed by Alexa 555-conjugated goat anti-rabbit antibody (red). Hoechst 33342 (blue) was used as a counterstain. GDNF staining is specific for neurons. Note that Schwann cells as well as satellite glial cells are both negative.
    • Anti-GDNF Antibody
      Expression of GDNF in rat hippocampus
      Immunohistochemical staining of rat hippocampus frozen sections with Anti-GDNF Antibody (#ANT-014), (1:100), (red). A. GDNF staining is apparent in the pyramidal layer (asterik), in astrocytic fibers (horizontal arrows) and in some interneurons (vertical arrow). B. Calbindin D28k (green) appears in a subset of cells in the pyramidal layer. C. Merge of GDNF and calbindin demonstrates some co-localization in neurons. DAPI is used as the counterstain (blue).
    Immunocytochemistry
    • Anti-GDNF Antibody
      Expression of GDNF in rat C6 glioma cells
      Immunocytochemical staining of GDNF in rat C6 glioma cells. A. Paraformaldehyde-fixed and permeabilized rat C6 glioma cells were stained with Anti-GDNF Antibody (#ANT-014), (1:200) followed by goat anti-rabbit-AlexaFluor-555 secondary antibody (red). Nuclei were visualized with the cell-permeable dye Hoechst 33342 (blue). B. Live view of the same field as in (A).
    References
    1. Lin, L.F. et al. (1993) Science 260, 1130.
    2. Airaksinen, M.S. and Saarma, M. (2002) Nat. Rev. Neurosci. 3, 383.
    3. Sariola, H. and Saarma, M. (2003) J. Cell Sci. 116, 3855.
    4. Paratcha, G. et al. (2003) Cell 113, 867.
    Scientific background

    Glial cell-line derived neurotrophic factor (GDNF) is a neurotrophic factor originally identified for its ability to promote the survival of dopaminergic neurons in the midbrain.1

    GDNF is part of the neurotrophic factor family known as GDNF-family ligand (GFL), which includes neurturin (NRTN), artemin (ARTN), and persephin (PSPN).2,3 The GFL ligands are structurally related to the transforming growth factor-b (TGF-b) protein superfamily and contain seven cysteine residues with the same relative spacing as other members of this family.2

    Typical of neurotrophic factors, GDNF is produced in the form of a precursor, preproGDNF, which is then cleaved during secretion to become proGDNF, and later processed to the mature (homodimeric) GDNF form.2

    The GFLs convey their activity by binding to a multicomponent receptor that includes the RET receptor tyrosine kinase and a glycosyl phosphatidylinositol (GPI)-linked ligand-binding subunit known as GDNF family receptor α (GFRα).2,3 Four different GFRα subunits have been identified (GFRα 1-4) that determine the specificity of the GFRα-RET receptor complex. Thus, GFRα1 together with RET make up the high affinity receptor for GDNF.

    The GDNF and GFRα complex interaction with RET induces activation of the intracellular tyrosine kinase domain of the latter. Phosphorylated RET can then interact with several intracellular signaling cascades including MAP kinase, PI3K and PLCγ signaling pathways.2,3  

    Lately a RET-independent signaling pathway for GDNF has been shown to involve binding of GDNF and GFRα to the neural cell adhesion molecule (NCAM). Signaling of GDNF via the NCAM complex stimulates intracellular tyrosine kinases such as Fyn and FAK.4

    GDNF is synthesized and secreted by a wide variety of cell types in the central and peripheral nervous system where it exerts its neurotrophic effects in the development and maintenance of distinct sets of neurons. Accordingly, GDNF knockout mice die shortly after birth, reflecting the central role of GDNF in neurodevelopment.

    Remarkably, GDNF was found to be an essential morphogen for kidney formation as well as to have a role is spermatogenesis.3

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 24/01/2020

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of human GDNF. Anti-GDNF Antibody (#ANT-014) can be used in western blot, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize GDNF from human, mouse and rat samples.

    For research purposes only, not for human use
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