Anti-Ret (extracellular) Antibody

Proto-Oncogene Tyrosine-Protein Kinase Receptor Ret, Proto-Oncogene C-Ret, Cadherin Family Member 12
    Cat #: ANT-025
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide CEWRQGDGKGITR, corresponding to amino acid residues 541-553 of human Ret (Accession P07949). Extracellular, N-terminus.
    Accession (Uniprot) Number P07949
    Gene ID 5979
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; rat - 12/13 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Specificity This antibody will recognize the intact Ret receptor and the cleaved Ret extracellular cell-membrane anchored fragment. It won't recognize the cleaved Ret kinase fragment.
    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).
    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.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, lci, wb
    May also work in: ip
    Western blot
    Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
    1,2. Anti-Ret (extracellular) Antibody (#ANT-025), (1:200).
    3,4. Anti-Ret (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of Ret in rat DRG
    Immunohistochemical staining of rat dorsal root ganglia (DRG) frozen sections using Anti-Ret (extracellular) Antibody (#ANT-025), (1:50). Ret staining (red) is detected in DRG neurons. Hoechst 33342 (blue) is used to stain cell nuclei.
    Indirect flow cytometry
    Indirect flow cytometry analysis of live intact human THP-1 monocytic leukemia cell line:
    ___ Unstained cells + goat-anti-rabbit-AlexaFluor-647 secondary antibody.
    ___ Cells + Anti-Ret (extracellular) Antibody (#ANT-025), (1:20) + goat-anti-rabbit-AlexaFluor-647 secondary antibody.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of Ret in human MCF-7 breast adenocarcinoma cells
    Immunocytochemical staining of live intact human MCF-7 breast adenocarcinoma cells. A. Extracellular staining of cells with Anti-Ret (extracellular) Antibody (#ANT-025), (1:25), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live image of the cells. C. Merge of the two images.
    References
    1. Durbec, P. et al. (1996) Nature 381, 789.
    2. Cabrera J.R. et al. (2011) J. Biol. Chem. 286, 14628.
    3. Anders, J. et al. (2001) J. Biol. Chem276, 35808.
    4. Runeberg-Roos, P. and Saarma, M. (2007) Ann. Med39, 572.
    5. Knowles, P.P. et al. (2006) J. Biol. Chem. 281, 33577.
    Scientific background

    The RET proto-oncogene on human chromosome 10q11.2, encodes a receptor tyrosine kinase (RTK) activated by members of the glial cell line-derived neurotrophic factor (GDNF) ligand family (GDNF, neurturin, artemin, and persephin) in conjunction with a ligand-specific coreceptor (GFRα1-4)1. RET belongs to the cadherin superfamily, and it has been suggested that the ret gene is the result of a recombination between a gene encoding a tyrosine kinase receptor and a gene encoding an ancestral cadherin at an early stage of evolution2. The extracellular domain of RET comprises, as observed for other tyrosine kinase receptors, a cysteine-rich domain, but also four cadherin-like domains3.

    RET signaling is crucial for the development of the enteric nervous system. RET also regulates the development of sympathetic, parasympathetic, motor, and sensory neurons, and is necessary for the postnatal maintenance of dopaminergic neurons. Outside the nervous system, RET is crucial for development of the kidney and plays a key role in spermatogenesis3.

    RET has attracted considerable clinical interest because of the range of mutations found in diverse conditions that include Hirschsprung disease and a variety of cancers involving the thyroid gland. RET-related cancers with thyroid involvement include sporadic and familial medullary thyroid carcinoma (MTC), multiple endocrine neoplasia 2 (MEN2) syndromes MEN2A and MEN2B, and papillary thyroid carcinoma (PTC)4.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, 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: 30/10/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an extracellular epitope located of human Ret. Anti-Ret (extracellular) Antibody (# ANT-025) can be used in western blot, indirect flow cytometry, immunohistochemistry and immunocytochemistry applications and has been designed to recognize Ret from rat, mouse and human samples.

    For research purposes only, not for human use