Anti-Human Orai1 (extracellula

Anti-Human Orai1 (extracellular)-FITC

CRACM1, TMEM142A
Cat #: ACC-060-F
Sizes: 50 µl
Source: Rabbit
Type: Polyclonal
Applications: FC
Reactivity: H

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 new version of the Anti-Human Orai1 (extracellular) antibody (#ACC-060) directly conjugated to FITC. The Anti-Human Orai1 (extracellular)-FITC antibody (#ACC-060-F) can be used in immunofluorescent applications such as direct flow cytometry and  recognizes Orai1 from human samples.

• Applications
• Specifications
• Scientific Background
• Related Products

Direct flow cytometry

Direct flow cytometry analysis of live Jurkat (human acute T cell leukemia) cells:
^___ Unstained cells.
^___ Cells + Anti-Human Orai1 (extracellular)-FITC antibody (#ACC-060-F), (10 µg/1x10⁶ cells).
The control antigen is not suitable for this application.
Immunogen
Peptide (C)KKQPGQPRPTSK, corresponding to amino acid residues 203-214 of human Orai1 (Accession Q96D31). 2^nd extracellular loop.

HomologyCanis - identical; bovine - 12/13 amino acid residues.
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 mouse or rat Orai1.
LabelFluorescein isothiocyanate (FITC).
Standard quality control of each lotWestern blot analysis (unlabeled antibody, #ACC-060), and direct flow cytometry analysis (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 The mechanism controlling intracellular Ca²⁺ level influx either by 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 identified.³ Orai1 (also known as CRACM1) acts as the store-operated calcium channel (SOC) and STIM1 acts as an endoplasmic reticulum (ER) Ca²⁺ sensor.^3,4 The formation of functional channels requires the presence of both Orai1 and STIM1 proteins working as a complex that involves co-clustering of Orai1 on the plasma membrane with STIM1 on the ER.^4-6TRPC1, a member of the transient receptor potential family was also suggested to act as a player in the SOC complex.⁷ 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-termini. In addition, two mammalian homologs of Orai1 have been identified: Orai2 and Orai3.3,9 All three (Orai1, Orai2, and Orai3) have been shown to be capable of forming store-operated channels.^9,10

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.
10. Lis, A. et al. (2007) Curr. Biol. 17, 794.