Anti-Two Pore Calcium Channel Protein 2 Antibody

TPCN2, TPC2
    Cat #: ACC-072
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide KKTLKSIRW(S)LPE(C) , corresponding to amino acid residues 187-199 of mouse Two pore calcium channel protein 2  with replacement of amino acid 192 with serine (S) (Accession Q8BWC0). 2nd luminal loop.
    Accession (Uniprot) Number Q8BWC0
    Gene ID 233979
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, rat, human - 12/13 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 -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of mouse kidney lysate (lanes 1 and 4), rat lung membrane (lanes 2 and 5) and human embryonic Kidney 293 cell lysate (lanes 3 and 6):
    1-3. Anti-Two Pore Calcium Channel Protein 2 Antibody (#ACC-072), (1:200).
    4-6. Anti-Two Pore Calcium Channel Protein 2 Antibody, preincubated with the control peptide antigen.
    Rat pulmonary arteries (1:2500) (Jiang, Y.L. et al. (2013) J. Biol. Chem. 288, 10381.).
    Immunohistochemistry
    Expression of Two pore calcium channel protein 2 in rat cerebellum
    Immunohistochemical staining of Two pore calcium channel protein 2 in rat cerebellum using Anti-Two Pore Calcium Channel Protein 2 Antibody (#ACC-072). A. TPC2 positive cells (green) appear in the granule cell layer (G) of the cerebellum (arrows). B. The extent of the granule layer is demonstrated with the counterstain DAPI (blue).
    References
    1. Zhu, M.X. et al. (2010) Am. J. Physiol. 298, C430.
    2. Ishibashi, K. et al. (2000) Biochem. Biophys. Res. Commun. 270, 370.
    3. Galione, A. et al. (2009) Pflugers Arch. 458, 869.
    4. Berridge, M.J. et al. (2000) Nat. Rev. Mol. Cell Biol. 1, 11.
    5. Lee, H.C. (2005) J. Biol. Chem. 280, 33693.
    6. Churchill, G.C. et al. (2002) Cell 111, 703.
    7. Brailoiu, E. et al. (2009) J. Cell. Biol. 186, 201.
    8. Calcraft, P.J. et al. (2009) Nature 459, 596.
    9. Ruas, M. et al. (2010) Curr. Biol. 20, 703.
    Scientific background

    Among various vertebrate species, three genes are known to encode two-pore segment channels (TPCs) termed TPC1-3. Interestingly TPC3 seems to be absent from the genomes of primates and rodents1. The primary sequence of these channels indicates the presence of two putative pore-forming repeats. Each repeat contains six transmembrane domains and a pore loop, a structure strikingly reminiscent of many voltage-gated Na+ (NaV) and Ca2+ (CaV) channels2. These twelve transmembrane structures are further thought to form functional dimers3. Both TPC1 and TPC2 show ubiquitous expression, while that of TPC1 is exceptionally high in spleen, lung, liver, and kidney2.

    Ca2+-mobilizing messengers such as inositol triphosphate, cyclic ADP ribose and nicotinic acid adenosine dinucleotide phosphate (NAADP) are responsible for the intracellular changes in Ca2+ ion concentration4.

    In contrast to the other Ca2+-mobilizing agents, NAADP, the most potent of these Ca2+ releasing molecules increases the cytosolic Ca2+ concentration via Ca2+ channels located on acidic vesicles (endolysosomes)5,6. Only quite recently, after almost a decade of being cloned, TPC1 and TPC2 were both found to be responsible for the NAADP-induced release of Ca2+ 7,8. Evidence that these two channels are indeed responsible for the release of Ca2+ is quite compelling since overexpression of TPC1 and its knockdown or point mutation of a critical residue increase and exacerbate Ca2+ release respectively7. In addition, b-cells from TPC2 knockout mice exhibited no Ca2+ release from endolysosomes upon NAADP stimulation8. Finally, in a study using immunopurified channels, it was demonstrated that TPC1 and TPC2 both respond to very low concentrations of NAADP and are unequivocally responsible for the release of Ca2+, whereas TPC3 may negatively regulate the release of Ca2+ 9.

    As these channels have only recently been discovered, very little is known about their physiology and gating mechanisms. Their probable involvement in a number of diseases such as lysosomal storage disease (LSDs), caused by the dysfunction of lysosomal associated proteins, has yet to be deciphered1.

    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: 25/11/2018

    Anti-Two Pore Calcium Channel Protein 2 Antibody (#ACC-072) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize TPC2 from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    More product citations
    1. Jiang, Y.L. et al. (2013) J. Biol. Chem. 288, 10381.
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