Anti-Orai1 Antibody

Product Name

Anti-Orai1 Antibody

View all ORAI1 Antibodies

SKU/Catalog Number

A00909

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Orai1 Antibody catalog # A00909. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Orai1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00909)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human Orai1 recombinant protein (Position: A49-A301).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00909 is reactive to ORAI1 in Human, Mouse, Rat

Applications

A00909 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

39 kDa

Calculated molecular weight

Background of ORAI1

ORAI1 (ORAI calcium release-activated calcium modulator 1), also known as CRACM1, TMEM142A, Calcium release-activated calcium channel protein 1, Protein orai-1, Transmembrane protein 142A, FLJ14466, is a calcium selectiveion channelthat in humans is encoded by the ORAI1 gene. Orai1 channels play an important role in the activation of T-lymphocytes. The loss of function mutation of Orai1 causes severe combined immunodeficiency (SCID) in humans. The mammalian orai family has two additional homologs, orai2 and orai3. Orai proteins share no homology with any other ion channel family of any other known proteins. They have 4 transmembrane domains and form tetramers.Prakriya et al. showed that ORAI1 is a PM protein, and that CRAC channel function is sensitive to mutation of 2 conserved acidic residues in the transmembrane segments. Glu106-to-asp (E106D) and glu190-to-gln (E190Q) substitutions in transmembrane helices 1 and 3, respectively, diminished calcium ion influx, increased current carried by monovalent cations, and rendered the channel permeable to cesium ion.Prakriya et al. showed that ORAI1 is a PM protein, and that CRAC channel function is sensitive to mutation of 2 conserved acidic residues in the transmembrane segments.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconstitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml
Flow Cytometry (Fixed), 1-3μg/1x10⁶ cells
Direct ELISA, 0.1-0.5μg/ml

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of ORAI1 using anti-ORAI1 antibody (A00909).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human A375 whole cell lysates,
Lane 2: human Jurka whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: human HepG2 whole cell lysates,
Lane 6: human K562 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ORAI1 antigen affinity purified polyclonal antibody (Catalog # A00909) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ORAI1 at approximately 39KD. The expected band size for ORAI1 is at 33KD.

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Figure 2. Flow Cytometry analysis of CACO-2 cells using anti-ORAI1 antibody (A00909).
Overlay histogram showing CACO-2 cells stained with A00909 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ORAI1 Antibody (A00909,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 3. Flow Cytometry analysis of A549 cells using anti-ORAI1 antibody (A00909).
Overlay histogram showing A549 cells stained with A00909 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ORAI1 Antibody (A00909,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For ORAI1 (Source: Uniprot.org, NCBI)

Alternative Names

CRACM1; CRACM1ORAT1; FLJ14466; ORAI calcium release-activated calcium modulator 1; Orai1; ORAT1; Protein orai-1; TMEM142A; TMEM142Acalcium release-activated calcium modulator 1; Transmembrane protein 142Acalcium release-activated calcium channel protein 1 ORAI1 CRACM1, IMD9, ORAT1, TAM2, TMEM142A ORAI calcium release-activated calcium modulator 1 calcium release-activated calcium channel protein 1|calcium release-activated calcium modulator 1|protein orai-1|transmembrane protein 142A

For more info on ORAI1, check out the ORAI1 Infographic

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In this infographic, you will see the following information for ORAI1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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