Anti-TRPV4 Antibody

OTRPC4, VRL-2, VR-OAC, TRP12
    Cat #: ACC-034
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide CDGHQQGYAPKWRAEDAPL, corresponding to amino acid residues 853-871 of rat TRPV4 (Accession Q9ERZ8). Intracellular, C-terminus.
    Accession (Uniprot) Number Q9ERZ8
    Gene ID 66026
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 17/19 amino acid residues identical; human - 16/19 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 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 -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, ip, wb
    Western blot
    Western blot analysis of ND7/23 cell line lysate:
    1. Anti-TRPV4 Antibody (#ACC-034), (1:200).
    2. Anti-TRPV4 Antibody, preincubated with the control peptide antigen.
    Western blot analysis of rat brain lysates:
    1. Anti-TRPV4 Antibody (#ACC-034), (1:200).
    2. Anti-TRPV4 Antibody, preincubated with the control peptide antigen.
    Immunoprecipitation
    Rat and mouse brain lysates (10 μg Ab/0.500 mg whole-protein) (Benfenati, V. et al. (2011) Proc. Natl. Acad. Sci. U.S.A. 108, 2563).
    Immunohistochemistry
    Expression of TRPV4 in rat DRG
    Immunohistochemical staining of rat dorsal root ganglion (DRG) frozen sections using Anti-TRPV4 Antibody (#ACC-034). A. TRPV4 (red) in DRG neurons. B. Staining with mouse anti-Parvalbumin (green) in the same DRG section. C. Confocal merge of TRPV4 and Parvalbumin demonstrates colocalization.
    Immunocytochemistry
    Expression of TRPV4 in rat DRG primary culture
    Immunocytochemical staining of paraformaldehyde-fixed and permeabilized rat dorsal root ganglion (DRG) primary culture.
    A, D. Staining using Anti-TRPV4 Antibody (#ACC-034), (1:500), followed by goat anti-rabbit-AlexaFluor-555 secondary antibody.
    B, E. Nuclear staining of cells using the cell-permeable dye Hoechst 33342.
    C. Merged image of panels A and B.
    F. Merged image of panels D and E.
    Indirect flow cytometry
    Human corneal epithelial cells (HCEC) (1:200) (Pan, Z. et al. (2008) Cell Calcium 44, 374.).
    The control antigen is not suitable for this application.
    References
    1. Montell, Cet al. (2002) Mol. Cell9, 229.
    2. Clapham, D.E. (2003) Nature 426, 517.
    3. Moran, M.M. et al. (2004) Curr.Opin.Neurobiol14, 362.
    4. Clapham, D.Eet al. (2003) Pharmacol. Rev55, 591.
    5. Gunthorpe, M.J.et al. (2002) Trends. Pharmacol. Sci. 23, 183.
    6. Peng, J.Bet al. (2003) J. Physiol551.3, 729.
    Scientific background

    TRP channels are a large family (about 28 genes) of plasma membrane, non-selective cationic channels that are either specifically or ubiquitously expressed in excitable and non-excitable cells.1 According to IUPHAR the TRP family comprises of three main subfamilies on the basis of sequence homology; TRPC, TRPM and TRPV (to date, three extra subfamilies are considered to belong to the TRP family; the TRPA, TRPML, and TRPP).1-4 The TRPV subfamily consists of six members, TRPV1-6.5

    TRPV4 (also named OTRPC4) is activated under hypotonic conditions and serves as an osmoreceptor. TRPV4 is expressed in brain, liver, kidney, heart, testis and salivary gland.6

    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:


    Knockout validation of Anti-TRPV4 Antibody in mouse kidney.Immunohistochemical staining of mouse kidney sections using Anti-TRPV4 Antibody (#ACC-034) and Anti-Aquaporin 2-ATTO-550 Antibody (#AQP-002-AO). TRPV4 staining (green, upper panels) is detected in both the connecting tubules (CNT) and the collecting ducts (CCD). AQP2 immunostaining (red) is also observed in the CCD and CNT. TRPV4 staining is not observed in the sections from TRPV4-/- mice. Adapted from Berrout, J. et al. (2012) J. Biol. Chem. 287, 8782. with permission of American Society for Biochemistry and Molecular Biology.

    Last update: 20/11/2018

    Anti-TRPV4 Antibody (#ACC-034) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunoprecipitation, indirect flow cytometry, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize TRPV4 from human, rat, and mouse samples.

    For research purposes only, not for human use
    Citations
    Published figures using this product
    Expression of TRPV4 in rat perihematomal area following ICH.
    Immunohistochemical staining of rat brain sections 24 hours after intracerebral hemorrhage (ICH) induction using Anti-TRPV4 Antibody (#ACC-034). At 24 hours post-ICH, TRPV4 immunostaining (red) was detected on neurovascular structures, perivascular astrocytes and endothelial cells in the perihematomal area. TRPV4 staining coincided with that of GFAP (lower panels), an astrocyte marker and von Willebrand factor (vWF) (upper panels), a marker of BBB. Nuclei were stained using DAPI.
    Adapted from Zhao, H. et al. (2018) Front. Mol. Neurosci. 11, 97. with permission of Frontiers.
    Immuno-colocalization of TRPV4 and KCa3.1 in human BSM cells.
    Immunocytochemical staining of human bronchial smooth muscle (HBSM) cells using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051), (green) and Anti-TRPV4 Antibody (#ACC-034), (red).
    Adapted from Yu, Z. et al. (2017) Front. Pharmacol. 8, 559. with permission of Frontiers.
    Downregulation of TRPV4 in rat retinal microvascular endothelium.
    Immunohistochemical staining of rat retinal sections using Anti-TRPV4 Antibody (#ACC-034). A. TRPV4 expression (green) is mostly detected in the endothelium of retinal microvessels. B. Specificity of the antibody is depicted by showing loss of TRPV4 staining when pre-incubating with the control peptide antigen. C. Significant decrease in TRPV4 expression is observed in retina section from diabetic animals.
    Adapted from Monaghan, K. et al. (2015) with kind permission of Curtis, T.M., Centre for Experimental Medicine, Queen’s University of Belfast.
    Immuno-colocalization of Aquaporin 2 and TRPV4 in rat kidney.
    Immunohistochemical staining of rat kidney sections using Anti-TRPV4 Antibody (#ACC-034) and Anti-Aquaporin 2-ATTO-550 Antibody (AQP-002-AO). Shown are Representative confocal plane micrographs (axes are shown) and corresponding cross-sections (pointed by arrows) showing three-dimensional stacks of TRPV4 (green), AQP2 localization (red), and the combined image localization (yellow). A. Wild-type Sprague-Dawley (S/D) rat. B. Cyst monolayer from a PCK453 rat. DAPI staining is shown in blue.
    Adapted from Zaika, O. et al. (2013) J. Am. Soc. Nephrol. 24, 604. with permission of the American Society of Nephrology.
    PKA activation causes TRPV4 subcellular translocation.
    Immunocytochemical staining of TRPV4 expression in split-opened nephrons using Anti-TRPV4 Antibody (#ACC-034). A. control conditions. B. Activation of PKA with forskolin leads to TRPV4 translocation to the apical plasma membrane. C. Activation of PKC with PMA has no effect. D. pre-treatment with PKA specific inhibitor, abolishes TRPV4 translocation.
    Adapted from Mamenko, M. et al. (2013) with permission of American Society for Biochemistry and Molecular Biology.
    KO validation citations
    1. Immunohistochemical staining of mouse kidney sections. Also tested in TRPV4-/- mice.
      Berrout, J. et al. (2012) J. Biol. Chem. 287, 8782.
    Western blot citations
    1. Rat microvascular endothelial cells.
      Suresh, K. et al. (2018) Am. J. Physiol. 314, L893.
    2. Mouse heart lysates.
      Dong, Q. et al. (2017) Sci. Rep. 7, 42678.
    3. Rat cerebral arterial myocyte lysates.
      Gebremedhin, D. et al. (2017) PLoS ONE 12, e0176796.
    4. Rat neonatal ventricle myocyte lysates.
      Wu, Q.F. et al. (2017) Cell Death Dis. 8, e2828.
    5. Mouse CCDcl1 cell lysate (1:1000).
      Li, Y. et al. (2016) PLoS ONE 11, e0155006.
    6. Mouse DRG lysate (1:5000).
      Kim, S. et al. (2016) Sci. Signal. 9, Ra71.
    7. Mouse mesenteric arteries lysate.
      Yap, F.C. et al. (2016) Am. J. Physiol. 310, H1151.
    8. Mouse bone marrow-derived macrophage lysate.
      Scheraga, R.G. et al. (2016) J. Immunol. 196, 428.
    9. Human microvascular endothelial cell (HMEC-1) lysate (1:2000).
      Goedicke-Fritz, S. et al. (2015) Eur. J. Cell Biol. 94, 391.
    10. Mouse microvascular endothelial cell lysate (1:500).
      Suresh, K. et al. (2015) Am. J. Physiol. 309, L1467.
    11. Rat myometrial protein lysate (1:200).
      Ying, L. et al. (2015) Sci. Transl. Med. 7, 319ra204.
    12. Mouse adipose tissue lysate.
      Castellani, L. et al. (2014) J. Appl. Physiol. 116, 1272.
    13. Human preadipocytes.
      Che, H. et al. (2014) Pflugers Arch. 466, 947.
    14. RPMVEC cell lysate.
      Parker, J.C. et al. (2013) Physiol. Rep. 1, e00121.
    Immunoprecipitation citations
    1. Rat cortical collecting duct cells.
      Pizzoni, A. et al. (2018) J. Cell Biochem. 119, 4120.
    2. Mouse DRG lysates.
      Kim, S. et al. (2016) Sci. Signal. 9, Ra71.
    3. Rat and mouse brain lysates (10 µg/500 mg lysate).
      Benfenati, Vet al. (2011) Proc. Natl. Acad. Sci. U.S.A. 108, 2563.
    Immunohistochemistry citations
    1. Rat brain sections.
      Zhao, H. et al. (2018) Front. Mol. Neurosci. 11, 97.
    2. Mouse heart sections.
      Dong, Q. et al. (2017) Sci. Rep. 7, 42678.
    3. Rat cerebral arterial segment sections.
      Gebremedhin, D. et al. (2017) PLoS ONE 12, e0176796.
    4. Mouse cornea sections (1:200).
      Okada, Y. et al. (2016) PLoS ONE 11, e0167200.
    5. Mouse kidney sections (1:200).
      Li, Y. et al. (2016) PLoS ONE 11, e0155006.
    6. Mouse mesenteric arteries sections.
      Yap, F.C. et al. (2016) Am. J. Physiol. 310, H1151.
    7. Rat trigeminal sections (1:100).
      Meng, Q. et al. (2015) Am. J. Physiol. 309, C1.
    8. Rat retinal sections.
      Monaghan, K. et al. (2015) PLoS ONE 10, e0128359.
    9. Rat kidney sections (1:500).
      Zaika, O. et al. (2013) J. Am. Soc. Nephrol. 24, 604.
    10. Mouse kidney sections. Also tested in TRPV4-/- mice.
      Berrout, J. et al. (2012) J. Biol. Chem. 287, 8782.

    Immunocytochemistry citations
    1. Rat cerebral arterial myocytes.
      Gebremedhin, D. et al. (2017) PLoS ONE 12, e0176796.
    2. Human bronchial smooth muscle cells.
      Yu, Z. et al. (2017) Front. Pharmacol. 8, 559.
    3. Mouse CCDcl1 cells (1:200).
      Li, Y. et al. (2016) PLoS ONE 11, e0155006.
    4. Rat myometrial smooth muscle cells (1:200).
      Ying, L. et al. (2015) Sci. Transl. Med. 7, 319ra204.
    5. Rat pulmonary microvascular endothelial cells (RPMVEC) (1:200).
      Parker, J.C. et al. (2013) Physiol. Rep. 1, e00121.
    6. Mouse distal nephrons (1:1000).
      Mamenko, M. et al. (2013) J. Biol. Chem. 288, 20306.
    Indirect flow cytometry citations
    1. Human corneal epithelial cells (HCEC) (1:200).
      Pan, Z. et al. (2008) Cell Calcium 44, 374.
    More product citations
    1. Wen, L. et al. (2018) BMC Ophthalmology 18, 38.
    2. Zhao, H. et al. (2018) Front. Mol. Neurosci. 11, 97.
    3. Jo, A.O. et al. (2016) Proc. Natl. Acad. Sci. U.S.A. 113, 3885.
    4. Alexander, R. et al. (2013) Br. J. Pharmacol. 168, 761.
    5. Fawcett, K.Aet al. (2012) J. Neurol. Neurosurg. Psychiatry 83, 1204.
    6. Ye, Let al. (2012) Cell 151, 96.
    Related Products
      • Products for control experiments
        1. Anti-TRPV4 (extracellular) Antibody (#ACC-124). This antibody recognizes a different epitope and thus can also be used as a control for detecting TRPV4.
      • Antibodies for immuno-colocalization experiments
        1. Anti-TRPV4-ATTO-550 Antibody (#ACC-034-AO). A fluorescent labeled primary antibody. It can be used in immuno-colocalization studies in conjunction with any of our antibodies raised in rabbit.
      • Antibodies for live cell imaging experiments
        1. Anti-TRPV4 (extracellular) Antibody (#ACC-124). This antibody recognizes an extracellular epitope and can be used to detect TRPV4 in live cells.