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Anti-STIM1 (extracellular) Antibody

Stromal interaction molecule 1, GOK
Cat #: ACC-063
Alternative Name Stromal interaction molecule 1, GOK
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide CHSEDEKLSFEAVR, corresponding to amino acid residues 56-69 of human STIM1 (Accession Q13586). STIM1 expressed on the plasma membrane: Extracellular, N-terminus. STIM1 expressed on the ER: Luminal, N-terminus.
Accession (Uniprot) Number Q13586
Gene ID 6786
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat, mouse, bovine, canis - identical.
RRID AB_2039893.
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.
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 0.45 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, ip, lci, wb
Western blot
  • Western blot analysis of RBL (lanes 1 and 4), HL-60 (lanes 2 and 5), and Jurkat (lanes 3 and 6) cell lysates:
    Western blot analysis of RBL (lanes 1 and 4), HL-60 (lanes 2 and 5), and Jurkat (lanes 3 and 6) cell lysates:
    1,2,3. Anti-STIM1 (extracellular) Antibody (#ACC-063), (1:1000).
    4,5,6. Anti-STIM1 (extracellular) Antibody, preincubated with STIM1 (extracellular) Blocking Peptide (#BLP-CC063).
Immunoprecipitation
  • Rat isolated cardiomyocytes (Saliba, Y. et al. (2015) Cardiovasc. Res. 105, 248.).
Immunohistochemistry
  • Expression of STIM1 in rat stomach
    Expression of STIM1 in rat stomach
    Immunohistochemical staining of paraffin embedded rat stomach sections using Anti-STIM1 (extracellular) Antibody (#ACC-063), (1:100). STIM1 is expressed in the parietal cells of the gastric mucosa (arrows). Hematoxilin is used as the counterstain.
Indirect flow cytometry
  • Cell surface detection of STIM1 in intact RBL (A) and Jurkat (B) cells.
    Cell surface detection of STIM1 in intact RBL (A) and Jurkat (B) cells.
    ___ Cells + FITC-conjugated goat anti-rabbit antibody.
    ___ Cells + Anti-STIM1 (extracellular) Antibody (#ACC-063), (5-10 µg antibody/0.5-1x106 cells) + FITC-conjugated goat anti-rabbit antibody.
  • The blocking peptide is not suitable for this application.
Live cell imaging / Immunocytochemistry
  • Expression of STIM1 in RBL cells
    Expression of STIM1 in RBL cells
    Cell surface detection of STIM1 in live rat basophilic leukemia (RBL) cells. A. Extracellular staining of cells with Anti-STIM1 (extracellular) Antibody (#ACC-063), (1:50) followed by goat anti-rabbit-AlexaFluor-555 secondary antibody. B. Nuclear staining of cells using the cell-permeable dye Hoechst 33342. C. Merged image of panels A and B.
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. Dziadek, M.A. and Johnstone, L.S. (2007) Cell Calcium 42, 123.
  6. Yuan, J.P. et al. (2007) Nat. Cell Biol. 9, 636.
  7. Muik, M. et al. (2008) J. Biol. Chem. 283, 8014.
  8. Li, Z. et al (2007) J. Biol. Chem. 282, 29448.
  9. Hewavitharana, T. et al. (2007) Cell Calcium 42, 173.
Scientific background

Cytosolic Ca2+ has long been known to act as a key second messenger in many intracellular pathways including synaptic transmission, muscle contraction, hormonal secretion, and cell growth and proliferation.1,2 The mechanism controlling the influx of intracellular Ca2+ either by calcium-release-activated Ca2+ channels (CRAC) or from intracellular stores has lately become of great interest.

Recently, several key players of the store-operated complex have been identified.The Orai family consists of three members, Orai1-3, and the STIM family, which consists of two members, STIM1 and STIM2. Orai1 (also known as CRACM1) acts as the store-operated calcium channel (SOC) and STIM1 as the endoplasmic reticulum (ER) Ca2+ sensor.3,4 The majority of STIM1 appears to be localized intracellularly at the ER membrane while low expression of STIM1 has been detected on the cell surface of several cell types.5 STIM1 has an amino-terminal EF hand Ca2+-binding domain facing the lumen of the ER.6 Upon Ca2+ store depletion, STIM1 molecules are redistributed in punctae underneath the plasma membrane and activate SOCs.

Several possible interactions between STIM1 and Orai1 have been suggested. The most simple and cited is a dynamic interaction between the cytosolic C-terminus of STIM1 and the cytoplasmic domain of the Orai1 channel.7-9 STIM1 is assumed to regulate the activity of all known SOCs, including native SOCs.Consistent with their important role as calcium sensors within the ER, STIM1 proteins are ubiquitously expressed.

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: 16/08/2020

Anti-STIM1 (extracellular) Antibody (#ACC-063) is a highly specific antibody directed against an epitope of the human Stromal interaction molecule 1. The antibody can be used in western blot, immunocytochemistry, immunohistochemistry, and indirect flow cytometry applications. It has been designed to recognize STIM1 from human, rat, and mouse samples.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Mouse NSC-34 motor neuron cell lysate (1:1000).
    Tedeschi Vet al. (2019) Sci Rep9, 10743.
  2. Rat lung lysates (1:1000).
    Zhang, W. et al. (2017) Inflammation 40, 778.
  3. Human HES2-CM cell lysate.
    Wang, Y. et al. (2015) Stem Cells 33, 2973.
  4. Rat isolated cardiomyocytes.
    Saliba, Y. et al. (2015) Cardiovasc. Res. 105, 248.
  5. Human primary coronary artery smooth cell lysate.
    Munoz, E. et al. (2013) Cell Calcium 54, 375.
Immunoprecipitation citations
  1. Mouse Motor Neuron (NSC-34) Cell Line (1:200).
    Tedeschi Vet al. (2019) Sci Rep9, 10743.
  2. Rat isolated cardiomyocytes (20 μg).
    Saliba, Y. et al. (2015) Cardiovasc. Res. 105, 248.
Immunohistochemistry citations
  1. Mouse brain sections (1:250).
    Kikuta, S. et al. (2019) Front. Cell. Neurosci. 13, 547.
  2. Rat lung sections (1:100).
    Zhang, W. et al. (2017) Inflammation 40, 778.
Immunocytochemistry citations
  1. Human HES2-Cm cells.
    Wang, Y. et al. (2015) Stem Cells 33, 2973.
Immunofluorescence citations
  1. Mouse brain sections (1:250).
    Kikuta, S. et al. (2019) Front. Cell. Neurosci. 13, 547.
More product citations
  1. Steinckwich, N. et al. (2011) J. Immunol. 186, 2182.
  2. Bomben, V.C. and Sontheimer, H. (2010) Glia 58, 1145.
  3. Bréchard, S. et al. (2009) Biochem. Pharmacol. 78, 504.
  4. Ohba, T. et al. (2009) Biochem. Biophys. Res. Commun. 389, 172.
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