Anti-RAMP1 (extracellular) Antibody

Receptor activity-modifying protein 1, Calcitonin Receptor-Like Receptor Activity Modifying Protein 1
    Cat #: ARR-021
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: m, r
    Immunogen
    Peptide CRDPDYGTLIQE, corresponding to amino acid residues 27-38 of rat RAMP1 (Accession Q9JJ74). Extracellular, N-terminus.
    Accession (Uniprot) Number Q9JJ74
    Gene ID 58965
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human - not recommended for human samples.
    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 Not recommended for human samples.
    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 0.85 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, lci, wb
    May also work in: ifc, ih, ip
    Western blot
    Western blot analysis of rat brain (lanes 1 and 4), mouse brain (lanes 2 and 5) and rat pancreas (lanes 3 and 6):
    1-3. Anti-RAMP1 (extracellular) Antibody (#ARR-021).
    4-6. Anti-RAMP1 (extracellular) Antibody, preincubated with the control peptide antigen.
     
    The Antibody was used at a concentration of 1:400 for brain lysates and 1:200 for pancreas lysates.
    Live cell imaging / Immunocytochemistry
    Expression of RAMP1 in rat PC12 cells
    Immunocytochemical staining of intact living rat pheochromocytoma (PC12) cells. A. Extracellular staining of cells using Anti-RAMP1 (extracellular) Antibody (#ARR-021), (1:50), (red). B. Live view of cells.
    References
    1. Kadmiel, M. et al. (2012) Adv. Exp. Med. Biol. 2012, 744, 49.
    2. Jacob, A. et al. (2012) Adv. Exp. Med. Biol. 744, 87.
    3. Conner, A.C. et al. (2004) Biochem. Soc. Trans. 32, 843.
    4. Li, M. et al. (2014) PLoS One 9, e102356.
    5. Shi, B. et al. (2014) Exp. Biol. Med. 239, 356.
    Scientific background

    Receptor activity modifying protein-1 (RAMP1) is part of a three-member group (that further includes RAMP2 and RAMP3 – each encoded by a different gene) of transmembrane proteins that interact with G-protein coupled receptors (GPCRs) such as the calcitonin receptor-like receptors (CLRs)1. This interaction leads to the formation of the receptor for calcitonin gene related peptide (CGRP) and is necessary for the binding of the peptide to the receptor. The binding of RAMP2 and RAMP3 to CLR, on the other hand, creates the receptor for adrenomedullin, a peptide functionally and structurally related to calcitonin1,2.

    RAMP1 (as well as the two other RAMPs) has a short intracellular domain and a single transmembrane domain. Its extracellular region contains four conserved cysteine residues that putatively form two disulfide bonds. This domain confers specificity to the receptor complex, whereas the transmembrane chain allows the association of the RAMPs with the GPCRs. In contrast to the other RAMPs, RAMP1 lacks glycosylation sites and therefore is unable to migrate to the cell surface independently. To reach the cell surface, it aggregates inside the cytoplasm and forms heterodimers with calcitonin3.

    RAMP1 is highly prominent in the neuronal tissues (especially the trigeminal ganglion, striatum, cortex, olfactory bulb, dorsal root ganglion, and the spinal cord) but abundant as well in the electrical conducting system of the heart, blood vessels, and related sites1,2,4.
    While not dramatically affected, RAMP1-KO mice are reported to have higher blood pressure and to suffer from repressed wound healing process, while overexpression of RAMP1 in animals leads to extravasation and vasodilation2. Interestingly, asthmatic symptoms in mice model of asthma were mitigated in mice that were RAMP1-deficient4. Furthermore mesenchymal stem cells overexpressing RAMP1 were shown to improve cardiac function and to reduce occurrences of over-thickened blood vessels5.

    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 epitope of rat RAMP1. AntiRAMP1 (extracellular) Antibody (#ARR-021) can be used in western blot and live cell imaging applications. The antibody recognizes an extracellular epitope and is thus ideal for detecting the protein in living cells. It has been designed to recognize RAMP1 from rat and mouse samples. The antibody does not recognize RAMP1 from human samples.

    For research purposes only, not for human use