Anti-Orai1 (extracellular)-ATTO-488 Antibody

TMEM142A, CRACM1, Calcium release-activated calcium channel protein 1
    Cat #: ACC-062-AG
    Alternative Name TMEM142A, CRACM1, Calcium release-activated calcium channel protein 1
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: m, r
    Immunogen
    • Peptide (C)KFLPLKRQAGQPS, corresponding to amino acid residues 200-212 of mouse Orai1 (Accession Q8BWG9). 2nd extracellular loop.
    • Anti-Orai1 (extracellular)-ATTO-488 Antibody
    Accession (Uniprot) Number Q8BWG9
    Gene ID 109305
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat - identical.
    RRID AB_11122852.
    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.
    Specificity Will not recognize human Orai1.
    Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
    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 1 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).
    Negative 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.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative 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 (unlabeled antibody, #ACC-062), and direct flow cytometry (labeled antibody).
    Applications: fc, ic, if, lci
    May also work in: ih*
    Direct flow cytometry
    • WEHI living cells (5-10 µg antibody/0.5-1x106 cells).
    Live cell imaging / Immunocytochemistry
    • Anti-Orai1 (extracellular)-ATTO-488 Antibody
      Expression of Orai1 in rat RBL cells
      Cell surface detection of Orai1 in intact living rat basophilic leukemia (RBL) cells using Anti-Orai1 (extracellular)-ATTO-488 Antibody (#ACC-062-AG), (1:100). A. Extracellular staining of RBL cells (green). B. Merge of A and live view of the cells.
    References
    1. Eisner, D.A. et al. (2005) Exp. Physiol. 90, 3.
    2. Chakrabarti, R. and Chakrabarti, R. (2006) J. Cell. Biochem. 99, 1503.
    3. Feske, S. et al. (2006) Nature 441, 179.
    4. Pickett, J. (2006) Nature Reviews Mol. Cell Biol. 7, 794.
    5. Luik, R.M. et al. (2006) J. Cell. Biol. 174, 815.
    6. Wu, M.M. et al. (2006) J. Cell. Biol. 174, 803.
    7. Ong, H.L. et al. (2007) J. Biol. Chem. 282, 9105.
    8. Luik, R.M and Lewis, R.S. (2007) Trends Mol. Med. 13, 103.
    9. DeHaven, W.I. et al. (2007) J. Biol. Chem. 282, 17548.
    Scientific background

    Cytosolic calcium (Ca2+) has long been known to act as a key second messenger in many intracellular pathways including synaptic transmission, muscle contraction, hormonal secretion, cell growth and proliferation1,2. Intracellular Ca2+ levels are controlled by either the influx of Ca2+ through the calcium-release-activated Ca2+ channels (CRAC), or from intracellular stores which gained much attention.

    Recently, several key players of the store operated complex have been identified3. Orai1 (also known as CRACM1) acts as the store operated Ca2+ channel (SOC) and STIM1, which acts as the endoplasmic reticulum Ca2+ sensor3,4. The formation of functional channels requires the presence of both Orai1 and STIM1 proteins working as a complex and involves the co-clustering of Orai1 on the plasma membrane with STIM1 on the endoplasmic reticulum4-6. TRPC1, a member of the transient receptor potential family was also suggested to act as a player in the SOC complex7. In T-cells, Ca2+ entry following activation by antigen-receptor engagement occurs solely through CRAC channels where Orai1 constitutes the pore forming subunit3,8.

    Orai1 is a plasma membrane protein with four potential transmembrane domains and intracellular N- and C-terminus. In addition, two mammalian homologs to Orai1 have been identified; Orai2 and Orai33,9. All three, Orai1 Orai2 and Orai3, are capable of forming store operated channels with different magnitudes9.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, 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: 16/03/2020

    Anti-Orai1 (extracellular) Antibody (#ACC-062) is a highly specific antibody directed against an extracellular epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry, live cell imaging, and indirect flow cytometry applications. It has been designed to recognize the Orai1 channel from mouse and rat samples.

    Anti-Orai1 (extracellular)-ATTO-488 Antibody (#ACC-062-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-Orai1 (extracellular)-ATTO-488 Antibody has been tested in live cell imaging applications and is especially suited for experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
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