Anti-Orai1 (extracellular)-ATT

Anti-Orai1 (extracellular)-ATTO-488

CRACM1, TMEM142A
Cat #: ACC-1062-AG
Sizes: 50 µl
Source: Rabbit
Type: Polyclonal
Applications: IC, LCI
May also work in: IH
Reactivity: M, R

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, IC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IH- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

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

Alomone Labs is pleased to offer a new version of antibody that 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. The Anti-Orai1 (extracellular)-ATTO-488 antibody (#ACC-062-AG) has been tested in live cell imaging applications and is specially suited for experiments requiring simultaneous labeling of different markers.

• Applications
• Specifications
• Scientific Background
• Related Products

Live cell imaging / Immunocytochemistry

Expression of Orai1 in rat RBL cells
Immunocytochemical staining of 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.
Immunogen
Peptide (C)KFLPLKRQAGQPS, corresponding to amino acid residues 200-212 of mouse Orai1 (Accession Q8BWG9). 2^nd extracellular loop.

HomologyRat - identical.
PurityAffinity purified on immobilized antigen.
FormulationLyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.
SpecificityWill not recognize human Orai1.
LabelATTO-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.
Standard quality control of each lotWestern blot analysis (unlabeled antibody, #ACC-062), and direct flow cytometry (labeled antibody).
Peptide confirmationConfirmed by amino acid analysis and mass spectrometry.
Storage before reconstitutionThe antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution50 µl double distilled water (DDW).
Antibody concentration after reconstitution1 mg/ml.
Storage after reconstitutionThe 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 reconstitutionLyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
Control antigen reconstitution100 µl double distilled water (DDW).
Control antigen storage after reconstitution-20ºC.
Preadsorption Control1 µg peptide per 1 µg antibody.
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. 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⁺sensor^3,4. 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 reticulum^4-6. 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 subunit^3,8.

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 Orai3^3,9. All three, Orai1 Orai2 and Orai3, are capable of forming store operated channels with different magnitudes⁹.

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.