Anti-CALHM1-ATTO-594 Antibody

Calcium Homeostasis Modulator 1, FAM26C
    Cat #: ACC-101-AR
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)EAMNHDLELGHTHG, corresponding to amino acid residues 252-265 of human calcium homeostasis modulator protein 1  (Accession Q8IU99).
    Accession (Uniprot) Number Q8IU99
    Gene ID 255022
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse - 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.
    Label ATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × 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 (unlabeled antibody, #ACC-101), and immunohistochemistry (labeled antibody).
    Applications: ic, ih
    Immunohistochemistry
    Expression of CALHM1 in rat hippocampus
    Immunohistochemical staining of rat hippocampus using Anti-CALHM1-ATTO-594 Antibody (#ACC-101-AR).  A. Staining of CALHM1 appears only in the hippocampal CA1 pyramidal layer.  B. Nuclear staining using DAPI as the counterstain.  C. Merge of A and B.
    Immuno-colocalization of GluN1 and CALHM1 in mouse hippocampal CA1 region
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDAR1 (GluN1) (extracellular) Antibody (#AGC-001), (1:200) and Anti-CALHM1-ATTO-594 Antibody (#ACC-101-AR), (1:60). A. Sections were stained with Anti-NMDAR1 (GluN1) (extracellular) Antibody, followed by goat-anti-rabbit-Cy2 (green). Staining reveals expression in neurons of the pyramidal layer (an arrow points at the layer). B. The same section was incubated with Anti-CALHM1-ATTO-594 Antibody, (red). C. Merge of the two images demonstrates colocalization of GluN1 and CALHM1 in pyramidal neurons.
    References
    1. Green, K.N. and LaFerla, F.M. (2008) Neuron 59, 190.
    2. Koppel, J. et al. (2011) Mol. Med. 17, 974.
    3. Dreses-Werringloer, U. et al. (2008) Cell 133, 1149.
    Scientific background

    A misbalance in Ca2+ homeostasis seems to be related to the development of Alzheimer’s disease (AD). Indeed Neuronal Ca2+ balance may affect the levels of proteins associated with AD, such as amyloid-beta (Ab) and tau1.

    Calcium homeostasis modulator 1 (CALHM1) is a cerebral Ca2+ channel component responsible for controlling intracellular Ca2+ levels and Ab metabolism2,3. CALHM1 is a three transmembrane glycoprotein, mostly localized to the endoplasmic reticulum (ER) although it can be detected at the plasma membrane. A functional channel is formed by multiple subunits which has some structural similarities to the NMDA receptor. It is expressed in all brain regions1.

    In cultured cells, overexpression of CALHM1 increases intracellular Ca2+ levels and reduces Ab accumulation, thereby reinforcing its implication in the development of AD. Indeed, a polymorphism in the gene was found to affect the onset of AD2.

    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: 29/10/2018

    Anti-CALHM1 Antibody (#ACC-101) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize CALHM1 from rat, mouse, and human samples.

    Anti-CALHM1-ATTO-594 Antibody (#ACC-101-AR) is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-CALHM1-ATTO-594 Antibody has been tested in immunohistochemistry applications and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use