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Anti-Human Orai1 (extracellular) Antibody

TMEM142A, CRACM1, Calcium release-activated calcium channel protein 1
Cat #: ACC-060
Alternative Name TMEM142A, CRACM1, Calcium release-activated calcium channel protein 1
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h
Immunogen
  • Peptide (C)KKQPGQPRPTSK, corresponding to amino acid residues 203-214 of human Orai1 (Accession Q96D31). 2nd extracellular loop.
Accession (Uniprot) Number Q96D31
Gene ID 84876
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Canis - identical; bovine - 12/13 amino acid residues identical.
RRID AB_2039844.
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 mouse or rat Orai1.
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 0.65 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).
Standard quality control of each lot Western blot analysis.
Applications: ic, if, ifc, ih, wb
May also work in: ip*, lci*
Western blot
  • Western blot analysis of Jurkat (lanes 1 and 2) and HL-60 (lanes 3 and 4) cell lysates:
    Western blot analysis of Jurkat (lanes 1 and 2) and HL-60 (lanes 3 and 4) cell lysates:
    1,3. Anti-Human Orai1 (extracellular) Antibody (#ACC-060), (1:200).    
    2,4. Anti-Human Orai1 (extracellular) Antibody, preincubated with the control peptide antigen.
Indirect flow cytometry
  • Cell surface detection of Orai1 in intact HL-60 (A) and Jurkat cells (B).
    Cell surface detection of Orai1 in intact HL-60 (A) and Jurkat cells (B).
    ___ Unstained cells.
    ___ Cells + Anti-Human Orai1 (extracellular) Antibody (#ACC-060), (5-10 µg antibody/0.5-1 x 106 cells).
  • The control antigen is not suitable for this application.
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. Hong 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 proliferation.1,2 The mechanism controlling intracellular Ca2+ level influx either by the calcium-release-activated Ca2+ channels (CRAC), or from intracellular stores, has become of great interest.

Recently, several key players of the store operated complex have been identified3Orai1 (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 reticulum4-6TRPC1, 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
Image & Title:

Anti-Human Orai1 (extracellular) Antibody
Expression of Orai1 in human kidney.Immunohistochemical staining of human kidney sections using Anti-Human Orai1 (extracellular) Antibody (#ACC-060). In paraffin-embedded kidney sections, Orai1 is localized to kidney tubules, with stronger staining in the proximal tubules (upper left panel). Antibody preabsorbed with the control antigen (supplied with the antibody) was used as control to show the specificity of the antibody (lower left panel). In frozen kidney sections, Orai1 staining (green) is observed in renal tubules (upper right panel). Smooth muscle actin staining is shown in red (lower right panel).Adapted from Zeng, B. et al. (2017) Nat. Commun. 8, 1920. with permission of SPRINGER NATURE.

Last update: 16/08/2020

Anti-Human Orai1 (extracellular) Antibody (#ACC-060) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, and indirect flow cytometry applications. It has been designed to recognize Orai1 from human samples only.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Human kidney lysate.
    Zeng, B. et al. (2017) Nat. Commun. 8, 1920.
  2. Human HK2 kidney cell lysate.
    Mai, X. et al. (2016) J. Am. Soc. Nephrol. 27, 3063.
  3. Human OUMS-27 chondrosarcoma cell lysate.
    Inayama, M. et al. (2015) Cell Calcium 57, 337.
  4. Human airway smooth muscle cell lysate (1:1000).
    Sathish, V. et al. (2015) Cell. Physiol. Biochem. 36, 1101.
  5. Human umbilical vein endothelial cell (HUVEC) lysate.
    Zhou, M.H. et al. (2014) J. Biol. Chem. 289, 29446.
  6. Human primary coronary artery smooth cell lysate.
    Munoz, E. et al. (2013) Cell Calcium 54, 375.
  7. Human HL-60 promyelocytic cell lysate.
    Brechard, S. et al. (2008) Cell Calcium 44, 492.
Immunohistochemistry citations
  1. Human kidney sections.
    Zeng, B. et al. (2017) Nat. Commun. 8, 1920.
Immunocytochemistry citations
  1. Human DU-145 prostate cells (1:200).
    Stagno, M.J. et al. (2017) Cell Physiol. Biochem. 42, 1366.
More product citations
  1. Motiani, R.K. et al. (2013) FASEB J. 27, 63.
  2. Abcejo, A.J. et al. (2012) PLoS ONE 7, e44343.
  3. Mitchell, C.B. et al. (2012) J. Neurochem. 122, 1155.
  4. Park, K.H. et al. (2011) Science Signaling 4, ra31.
  5. Meuchel, L.W. et al. (2011) Am. J. Physiol. 301, L91.
  6. Li, J. et al. (2011) Br. J. Pharmacol. 164, 382.
  7. Steinckwich, N. et al. (2011) J. Immunol. 186, 2182.
  8. Lur, G. et al. (2011) Biochem. J. 436, 231.
  9. Schaff, U.Y. et al. (2010) Blood 115, 657.
  10. Bisaillon, J.M. et al. (2010) Am. J. Physiol. 298, C993.
  11. Bomben, V.C. and Sontheimer, H. (2010) Glia 58, 1145.
  12. Fernandez-Rodriguez, S. et al. (2009) Cardiovasc. Res. 84, 470.
  13. Bréchaud, S. et al. (2008) Cell Calcium 44, 492.
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