Anti-CaV3.2 (CACNA1H) Antibody

Voltage-dependent T-type calcium channel subunit α1H
Cat #: ACC-025
Alternative Name Voltage-dependent T-type calcium channel subunit α1H
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
      • Peptide CHVEGPQERARVAHS, corresponding to amino acid residues 581-595 of rat CaV3.2 (Accession number Q9EQ60). Intracellular loop between domains D1 and D2.
    Accession (Uniprot) Number Q9EQ60
    Gene ID 114862
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, bovine and canis - 14/15 amino acid residues identical; human - 13/15 amino acid residues identical.
    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).
    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, wb
    May also work in: ifc
      • Western blot analysis of rat DRG lysates:
        1. Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025), (1:200).
        2. Anti-CaV3.2 (CACNA1H) Antibody, preincubated with the control peptide antigen.
        Human CaV3.2 transfected in HEK-293 cells (1:100) (Markandeya, Y.S. et al. (2011) J. Biol. Chem. 286, 2433.).
      • Rat brain lysates (5 μg) (Weiss, N. et al. (2012) J. Biol. Chem. 287, 2810.).
      • Expression of CaV3.2 in mouse cerebellum
        Immunohistochemical staining of mouse cerebellum frozen sections with Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025), (1:100). A. CaV3.2 appears adjacent to Purkinje cells and in fibers in the molecular layer (red). B. Staining of Purkinje cells with mouse anti-parvalbumin (PV, green). C. Merged image of panels A and B demonstrates presence of CaV3.2 adjacent to Purkinje cells.
        Expression of CaV3.2 in rat DRG
        Immunohistochemical staining of rat dorsal root ganglion (DRG) frozen sections with Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025), (1:50). Staining is specific for DRG. Note that neither glial cells nor axonal fibers are stained. Hoechst 33342 is used as the counterstain.
      • Expression of CaV3.2 in rat DRG primary culture
        Immunocytochemical staining of paraformaldehyde-fixed and permeabilized rat dorsal root ganglion (DRG) primary culture.
        A, D. Immunocytochemical staining using Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025), (1:200), followed by goat anti-rabbit-AlexaFluor-488 secondary antibody.
        B, E. Nuclear fluorescence staining of cells using the membrane-permeable DNA dye Hoechst 33342.
        C. Merged image of panels A and B.
        F. Merged image of panels D and E.
         Magnification:
        A-C: x20
        D-F: x100
      • T-type Ca2+ channels play an important role in many cellular processes such as hormone secretion, neurotransmitter release and cell differentiation. T-type channels are also known to participate in the pacemaker activities of the heart and neurons including thalamic neurons.1

        Three genes encoding T-type Ca2+ channels have been cloned and designated as CaV3.2 (CACNA1H), CaV3.1 (CACNA1G) and CaV3.3 (CACNA1I).1-3  

        The CaV3.2 channel is widely expressed in various tissues such as the brain, heart, liver and testis.

        Involvement of CaV3.2 Ca2+ channels in several pathologies has been described. Overexpression of the CaV3.2 channel was described in prostate cancer cells that are associated with more aggressiveness, invasiveness and poor prognosis.4 Several point mutations discovered in the CaV3.2 channel that affect gating of the channel were found to be associated with Childhood Absence Epilepsy.5 One year old mice, deficient in CaV3.2 channel, exhibited severe cardiac pathology, fibrosis, necrosis, lymphocyte infiltration and abnormal coronary function compared to wild-type mice.

        Recently, it has been demonstrated that T-type channels are expressed in DRG neurons and that small and medium-diameter primary afferent neurons in the dorsal horn expresses almost exclusively the CaV3.2 Ca2+channels. This might indicate a possible role for CaV3.2 in nociception.7

    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-Cav3.2 (CACNA1H) Antibody
    Expression of CaV3.2 in Rat Sural Nerve.Immunohistochemical staining of rat sural nerve sections using Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025). A. CaV3.2 staining (green) partially colocalizes with neurofilament 200. B. CaV3.2 staining (green) significantly colocalizes with calcitonin gene-related peptide (CGRP, red), a marker for nociceptive peptidergic fibers.Adapted from Chen, W. et al. (2018) Front. Mol. Neurosci. 11, 24. with permission of Frontiers.

    Last update: 03/06/2019

    Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunoprecipitation, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize CaV3.2 from rat, human, and mouse samples.

    For research purposes only, not for human use
    Citations
      • Expression of CaV3.2 in rat carotid body.
        Left: mRNA expression of different CaV channels in the carotid body. Right: Immunohistochemical staining of rat carotid body using Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025). CaV3.2 staining (red) is detected in glomus cells. Use of the control peptide antigen eliminated the signal obtained using the antibody.
        Adapted from Makarenko, V.V. et al. (2015) with permission of the American physiological society.
        Upregulation of CaV3.2 under hypoxia.
        Immunocytochemical staining of rat pulmonary artery smooth muscle cells (PASMCs) and rat pheochromocytoma cells (PC12) using Anti-CaV3.2 (CACNA1H) Antibody (#ACC-025) and Anti-CACNA1G (CaV3.1) Antibody (#ACC-021). Staining shows upregulation of CaV3.2 and not CaV3.1 in response to oxygen stress.
        Adapted from Sellak, H. et al. (2014) with permission of the American Physiological Society.
      • Western blot analysis of mouse brain lysate. Tested in CACNA1H-/- mice.
        Proft, J. et al. (2017) Sci. Rep. 7, 11513.
      • Mouse brain lysate. Also tested in CACNA1H-/- mice.
        Proft, J. et al. (2017) Sci. Rep. 7, 11513.
      • HEK 293 cell lysates (1:500).
        Makarenko, V.V. et al. (2016) J. Neurophysiol. 115, 345.
      • Human artery lysates (1:200).
        Harraz, O.F. et al. (2015) J. Gen. Physiol. 145, 405.
      • Rat DRG lysates (1:200).
        Makarenko, V.V. et al. (2015) Am. J. Physiol. 308, C-146.
      • Rat hippocampus lysate (1:1500).
        Sanchez-Aguilera, A. et al. (2014) J. Physiol. 592, 2845.
      • Mouse lung and brain lysate (1:200).
        Wan, J. et al. (2013) Am. J. Physiol. 305, L154.
      • Human CaV3.2 transfected in HEK-293 cells (1:100).
        Markandeya, Y.S. et al. (2011) J. Biol. Chem. 286, 2433.
      • Rat brain lysates (5 μg).
        Weiss, N. et al. (2012) J. Biol. Chem. 287, 2810.
      • Rat sural nerve sections.
        Chen, W. et al. (2018) Front. Mol. Neurosci. 11, 24.
      • Rat DRG sections (1:100).
        Liu, Z. et al. (2016) Mol. Pain 12, 1.
      • Rat retinal whole mounts (1:200).
        Fernandez, J.A. et al. (2015) Invest. Ophtalmol. Vis. Sci. 56, 5125.
      • Rat carotid sections (1:200).
        Makarenko, V.V. et al. (2015) Am. J. Physiol. 308, C-146.
      • Rat DRGs (1:200).
        Huang, D. et al. (2016) Biochem. Biophys. Res. Commun. 473, 693.
      • Human NCI-H295R transfected cells (1:500).
        Reimer, E.N. et al. (2016) Endocrinology 157, 3016.
      • Rat pulmonary artery smooth muscle cells (PASMCs) and rat pheochromocytoma cells (PC12), (1:100).
        Sellak, H. et al. (2014) Am. J. Physiol. 307, C648.
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