Anti-TRPA1 (extracellular) Antibody

Transient receptor potential cation channel subfamily A member 1, ANKTM1, Ankyrin-like with transmembrane domains protein 1, Transformation-sensitive protein p120
    Cat #: ACC-037
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)NSTGIINETSDHSE, corresponding to amino acid residues 747-760 of human TRPA1 (Accession O75762). 1st extracellular loop.
    Accession (Uniprot) Number O75762
    Gene ID 8989
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Bovine - identical; canis - 13/14 amino acid residues identical; rat - 12/14 amino acid residues identical; mouse - 9/14 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.
    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 1 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 -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ih, ip, lci, wb
    May also work in: ifc
    Western blot
    Western blot analysis of rat DRG (lanes 1,2), non-differentiated PC12 cells (lanes 3,5) and differentiated PC12 cells (lanes 4,6) lysates:
    1,3,4. Anti-TRPA1 (extracellular) Antibody (#ACC-037), (1:200).
    2,5,6. Anti-TRPA1 (extracellular) Antibody, preincubated with the control peptide antigen.
    Human urethra (1:2000) (Gratzke, C. et al. (2009) Eur. Urol55, 696.).
    Immunoprecipitation
    Immunoprecipitation of PC-12 lysates:
    1. PC-12 lysates.
    2. PC-12 lysates + Anti-TRPA1 (extracellular) Antibody (#ACC-037) (6.5 µg) + protein A beads.
    3. Anti-TRPA1 (extracellular) Antibody + protein A beads.
    Immunohistochemistry
    Expression of TRPA1 in rat DRG
    Immunohistochemical staining of rat dorsal root ganglion (DRG) using Anti-TRPA1 (extracellular) Antibody (#ACC-037) (1:200). A. TRPA1 staining (green) is detected in both cells (horizontal arrow) and processes (vertical arrow). B. Nuclear staining using DAPI as the counterstain. C. Merged image of A and B.
    Human urethra (1:500) (Gratzke, C. et al. (2009) Eur. Urol55, 696.).
    Live cell imaging / Immunocytochemistry
    Expression of TRPA1 in rat PC12 cells
    Immunocytochemical staining of intact living rat PC12 cells. A. Extracellular staining of cells using Anti-TRPA1 (extracellular) Antibody (#ACC-037), (1:50) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Merged image of A and B.
    References
    1. Hill, K. and Schaefer, M. (2007) J. Biol. Chem. 282, 7145
    2. Story, G.M. et al. (2003) Cell 112, 819.
    3. Bandell, M. et al. (2004) Neuron 41, 849.
    4. Jordt, S.E. et al. (2004) Nature 427, 260.
    5. Zurborg, S. et al. (2007) Nat. Neurosci. 10, 277.
    6. Corey, D.P. et al. (2004) Nature 432, 723.
    7. Sotomayor, M. et al. (2005) Structure 13, 669.
    8. Clapham, D.E. (2002) Science 295, 2228.
    Scientific background

    The TRPA family is comprised of only one mammalian member, the TRPA1 (formerly named ANKTM1). TRPA1 is expressed in peripheral sensory neurons, where it is suggested to contribute to the detection of painful stimuli.1

    Originally, it was thought that TRPA channels sensed painfully cold temperatures,2 but a more conservative description is that TRPA1 is sensitive to membrane/cytoskeletal perturbations caused by low temperatures3-5 and perhaps stretch.6 In addition, it is sensitive to pungent natural compounds present in cinnamon oil, mustard oil, and wintergreen oil.

    TRPA1 is also expressed in hair cells, where its role in sensing mechanical forces is still unclear and controversial.1

    TRPA1 has a similar structure to all other TRP ion channels; six transmembrane domains, intracellular N-and C-terminus. However, the N-terminal domain possesses 17 ankyrin repeats that might indicate its potential role as a mechanosensor.6,7

    In addition, TRPA1 is expressed in nociceptive neurons expressing TRPV1 and might serve as a marker for polymodal nociceptors.8

    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
    Image & Title: Anti-TRPA1 (extracellular) Antibody
    Expression of TRPA1 in rat DRGs.
    Immunohistochemical staining of rat DRG sections using Anti-TRPA1 (extracellular) Antibody (#ACC-037). TRPA1 staining (red) is detected in DRGs and co-localizes with IB4 staining (green).Adapted from Yamamoto, K. et al. (2015) Mol. Pain 11, 69. with permission of BioMed Central.
    Last update: 05/11/2018

    Anti-TRPA1 (extracellular) Antibody (#ACC-037) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunoprecipitation immunohistochemistry, and live cell imaging applications. It has been designed to recognize TRPA1 from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Rat DRG lysate (1:200).
      Yamamoto, K. et al. (2016) Mol. Pain 11, 69.
    2. Mouse cerebral and cerebellar arteries lysate (1:500). Also tested in TRPA1 knock out mice.
      Sullivan, M.N. et al. (2015) Sci. Signal. 8, 358.
    3. Human cerebral arteries.
      Sullivan, M.N. et al. (2015) Sci. Signal. 8, 358.
    4. Human urethra (1:2000).
      Gratzke, C. et al. (2009) Eur. Urol55, 696.
    Immunoprecipitation citations
    1. Mouse cerebral and cerebellar arteries lysate 2 μg. Also tested in TRPA1 knock out mice.
      Sullivan, M.N. et al. (2015) Sci. Signal. 8, 358.
    2. Human cerebral arteries.
      Sullivan, M.N. et al. (2015) Sci. Signal. 8, 358.
    Immunohistochemistry citations
    1. Rat trigeminal sections (1:100).
      Meng, Q. et al. (2015) Am. J. Physiol. 309, C1.
    2. Rat DRG sections.
      Yamamoto, K. et al. (2015) Mol. Pain 11, 69.
    3. Human urethra (1:2000).
      Gratzke, C. et al. (2009) Eur. Urol55, 696.
    More product citations
    1. Weinhold, P. et al. (2010) J. Urol. 183, 2070.
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