Anti-CACNA1B (CaV2.2) Antibody

Voltage-dependent N-type calcium channel subunit α1B, Brain calcium channel III, BIII
Cat #: ACC-002
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RHHRHRDRDKTSASTPA, corresponding to amino acid residues 851-867 of rat CACNA1B (Accession Q02294). Intracellular loop between domains II and III.
    Accession (Uniprot) Number Q02294
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 15/17 amino acid residues identical; human 11/17 amino acid residues identical; rabbit - 11/17 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, ih, ip, wb
    May also work in: ifc
    Western blot
    Western blot analysis of rat brain membranes:
    1. Anti-CACNA1B (CaV2.2) Antibody (#ACC-002), (1:200).
    2. Anti-CACNA1B (CaV2.2) Antibody, preincubated with the control antigen.
    Immunoprecipitation
    Transfected tsA201 cells (1:200) (Marangoudakis, S. et al. (2012) J. Neurosci. 32, 10365.).
    Immunohistochemistry
    Expression of CaV2.2 in mouse cerebellum
    Immunohistochemical staining of mouse cerebellum with Anti-CACNA1B (CaV2.2) Antibody (#ACC-002), (1:100). A. CaV2.2 (red) appears in Purkinje cells (arrows) and is distributed diffusely in the molecular layer (Mol). B. Staining of Purkinje cells with mouse anti-Calbindin 28K (green) demonstrates the restriction of CaV2.2 to cell bodies but not to dendrites in the molecular layer. C. Merged image of panels A and B.
    Immunocytochemistry
    Expression of CaV2.2 in rat DRG primary culture
    Immunocytochemical staining of paraformaldehyde-fixed and permeabilized rat dorsal root ganglion (DRG) primary culture. A. Cells were stained using Anti-CACNA1B (CaV2.2) Antibody (#ACC-002), (1:200) followed by goat anti-rabbit-AlexaFluor-555 secondary antibody. B. Nuclear fluorescence staining of cells using the membrane-permeable DNA dye Hoechst 33342. C. Merged images of panels A and B.
    NCI-H295R human adrenocortical cell line (H295R) (1:100) (Aritomi, S. et al. (2011) Hypertens. Res. 34, 193.).
    References
    1. Tsunemi, T. et al. (2002) J. Biol. Chem. 277, 7214.
    2. Beuckmann, C.T. et al. (2003) J. Neurosci. 23, 6793.
    3. Scheuber, A. et al. (2004) J. Neurosci. 24, 10402.
    4. Dascal, N. (2001) Trends Endocrinol. Metab. 12, 391.
    5. McCleskey, E.W. et al. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 4327.
    6. Feng, Z.P. et al. (2001) J. Biol. Chem. 276, 15728.
    Scientific background

    Voltage-dependent Ca2+ channels (CaV channels) are pivotal players in many physiological roles such as secretion, contraction, migration and excitation.1

    The voltage-dependent Ca2+ channels are composed of several subunits; α1, β, α2δ and γ. CaV channels were originally divided into six physiological types: L-, N-, P-, Q-, R-, and T-type.

    The CaV2.2 (formally named α1B) composes the α1 poreforming subunit for the N-type Ca2+ channel family. They are involved in neurotransmitter release from central neurons, including glutamate, γ-aminobutyric acid, acetylcholine, dopamine and noradrenaline.2

    The CaV2.2 is expressed preferentially in the central nervous system, where along with CaV2.1, it is responsible for pre-synaptic Ca2+ influx and neurotransmitter release.1,3

    The CaV2.2 channel is negatively regulated by many different GPCRs. There are two ways that this is done: either by directly binding Gβγ to the channel or by an indirect mechanism involving second messenger and channel phosphorylation.

    ω-Conotoxin GVIA (#C-300) is a specific blocker of CaV2.2 Ca2+ channels. It specifically blocks N-type CaV channels by binding to the CaV2.2 α1 subunit (α1B) and its action is only partially reversible.5,6

    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: Anti-CACNA1B (Cav2.2) AntibodyExpression of CaV α subunits in rat ICG.Immunohistochemical staining of rat intracardiac ganglia (ICG) using Anti-CaV1.2 (CACNA1C) Antibody (#ACC-003), Anti-CaV1.3 (CACNA1D) Antibody (#ACC-005), Anti-CACNA1A (CaV2.1) Antibody (#ACC-001), Anti-CACNA1B (CaV2.2) Antibody (#ACC-002) and Anti-CaV2.3 (CACNA1E) Antibody (#ACC-006). All CaV subtypes are expressed in sham treated rats but N-type CaV channel levels are decreased in CHF rats.Adapted from Tu, H. et al. (2014) with permission of the American Physiological Society.
    Last update: 21/11/2018

    Anti-CACNA1B (CaV2.2) Antibody (#ACC-002) 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 CaV2.2 from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse calyx of Held lysate.
      Sun, Z.C. et al. (2016) Sci. Rep. 6, 21774.
    2. Rat primary submucosa cells.
      Rehn, M. et al. (2013) Cell Tissue Res. 353, 355.
    3. Mouse synaptosome (1:300).
      Saggu, S. et al. (2013) Schizophr. Res. 146, 254.
    Immunoprecipitation citations
    1. Transfected tsA201 cells (1:200).
      Marangoudakis, S. et al. (2012) J. Neurosci. 32, 10365.
    Immunohistochemistry citations
    1. Mouse kidney sections.
      Ohno, S. et al. (2016) Sci. Rep. 6, 27192.
    2. Rat retinal sections (1:1000).
      Sargoy, A. et al. (2014) PLoS ONE 9, e84507.
    3. Rat intracardiac ganglia and stellate ganglia neurons.
      Tu, H. et al. (2014) Am. J. Physiol. 306, C132.
    4. Rat retinal sections (1:1000-1:1500).
      Liu, X. et al. (2013) J. Physiol. 591, 3309.
    Immunocytochemistry citations
    1. Rat adipose-derived stromal cells and rat bone marrow stromal cells.
      Forostyak, O. et al. (2016) Stem Cell Res. 16, 622.
    2. Rat stellate neurons (1:200).
      Larsen, H.E. et al. (2016) J. Neurosci. 36, 8562.
    3. Rat primary submucosa cells (1:100).
      Rehn, M. et al. (2013) Cell Tissue Res. 353, 355.
    4. NCI-H295R human adrenocortical cell line (H295R) (1:100).
      Aritomi, S. et al. (2011) Hypertens. Res. 34, 193.
    More product citations
    1. Mishima, K. et al. (2013) Am. J. Physiol. 304, F665.
    2. Lv, P. et al. (2012) J. Neurosci. 32, 16314.
    3. Shoham, S. et al. (2001) Biol. Psychiatry 49, 876.
    4. Schiff, M.L. et al. (2000) Nature 408, 723.
    5. Singer-Lahat, D. et al. (2000) Eur. J. Physiol. 440, 627.
    6. Bae, I.H. et al. (1999) Korean J. Biol. Sci. 3, 53.
    7. Barbosa, J. et al. (1999) J. Neurochem. 73, 1881.
    8. Lopez, I. et al. (1999) Neuroscience 92, 773.
    9. Macleod, G.T. et al. (1999) J. Neurophysiol. 82, 1133.
    10. Serrano, C.J. et al. (1999) FEBS Letters 462, 171.
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