This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ORAI2 / Protein orai-2 · Western blot design guide
Design a Western Blot for ORAI2
Source-linked ORAI2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ORAI2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
ORAI2 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~28.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Oligomerization-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ORAI2 Western Blot Protocol Options
The A07411 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog A07411) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A07411 · (A) 1, (B) 2 and (C) 4 μg/mL (catalog A07411) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected ORAI2 Western Blot Band Size?
ORAI2 has a predicted 28.6 kDa subunit mass; membrane localization and oligomerization could affect detection, but no empirical migration is supplied.
| Band near 28.6 kDa | Consistent with the predicted ORAI2 subunit size; confirm identity with controls |
| Weak or absent band in soluble lysate | ORAI2 is a multi-pass cell-membrane protein that may be poorly recovered |
| Band enriched in membrane fraction | Consistent with ORAI2 cell-membrane localization |
| Higher-mass band | Could reflect retained ORAI complexes if denaturation is incomplete; confirm identity |
💡Expected ORAI2 appearanceUniProt predicts a 28.6 kDa ORAI2 subunit, but no empirical band size is supplied; use membrane-enriched samples and band-identity controls to assess any detected band.
| UniProt predicted mass | Places the full-length ORAI2 subunit near 28.6 kDa |
| 254-amino-acid sequence | Defines the full-length protein underlying the predicted mass |
| Homomeric ORAI assembly | May yield higher-mass material if complexes persist during sample preparation; no apparent size is established |
| Heteromeric ORAI assembly | May yield higher-mass material if complexes persist during sample preparation; no apparent size is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ORAI2 may be poorly extracted | Check membrane-enriched material and extraction conditions |
| Band higher than expected | ORAI complexes may persist if denaturation is incomplete | Check denaturation conditions and verify band identity with ORAI2 depletion |
| Band lower than expected | The supplied features do not establish a smaller ORAI2 form | Check sample integrity and verify band identity with ORAI2 depletion |
| Multiple bands | ORAI complex retention or nonspecific antibody binding may contribute | Compare denaturation conditions and verify bands with ORAI2 depletion |
| Weak or no signal | Low recovery of multi-pass membrane ORAI2 is possible | Check membrane extraction and include a positive-control lysate |
Experiment evidence
Western blot images showing ORAI2 band
A07411 · ORAI2
A07411-1 · ORAI2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ORAI2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ORAI2 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: As a multi-pass membrane protein, ORAI2 may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for ORAI2
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Ovary | follicle cells | High | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ORAI2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ORAI2, answered from its protein features.
Where should the ORAI2 band appear?
Band shift · ORAI2 has a calculated mass of 28.6 kDa. No observed Western blot band position is supplied, so use 28.6 kDa as a reference, not a confirmed apparent mass.
Could an alternative isoform explain another band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning another band to an ORAI2 isoform.
Could a modification explain a band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not identify a modification that explains a shift, and they do not establish whether a shift occurs.
Does this guide establish induction of ORAI2?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for ORAI2 Western blot?
Transfer · ORAI2 is a 254-residue, multi-pass cell-membrane protein with a calculated mass of 28.6 kDa. Check that your transfer retains protein near this mass and confirm transfer on the membrane. The supplied features do not specify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07411 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should ORAI2 bands be quantified?
Quantitation · Quantify a consistently assigned ORAI2 band within the assay's linear range. If comparing store-depleted samples, interpret changes in band intensity separately from the reported STIM1 colocalization; that feature alone does not establish a change in total ORAI2.
Does store depletion increase ORAI2 expression?
Interpretation · The record says ORAI2 colocalizes with STIM1 upon store depletion. That supports a change in colocalization, but does not establish increased ORAI2 abundance.
ORAI2 can form homomeric and heteromeric ORAI complexes, and native CRAC channels are thought to contain hexameric ORAI heteromers. This makes complexes a possible interpretation to investigate, but the feature does not demonstrate that a higher Western blot band is an ORAI2 complex.
ORAI2 can form homomeric and heteromeric ORAI complexes, and native CRAC channels are thought to contain hexameric ORAI heteromers. This makes complexes a possible interpretation to investigate, but the feature does not demonstrate that a higher Western blot band is an ORAI2 complex.
Boster reagents
ORAI2 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Protein orai-2 ORAI2 Antibody
Cat # A07411
Real WB data
Anti-Orai2 Antibody Picoband®
Cat # A07411-1
Both catalogued anti-ORAI2 antibodies have Western blot images. A07411 was tested on Jurkat lysate at 1–4 μg/mL; A07411-1 shows an approximately 39 kDa band in four human cell lysates at 0.5 μg/mL. No publication or independent validation evidence was supplied.
Which to pick: For human samples, A07411-1 has WB images covering A375, Jurkat, HepG2, and K562 lysates; A07411 has a Jurkat WB image. For mouse work, A07411 lists mouse reactivity, but its supplied WB image uses Jurkat lysate.
Source: BosterBio ORAI2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96SN7.
- Human Protein Atlas. ORAI2 tissue expression.
- PMC5355949 — target-verified WB comparison
- PMC3121304 — target-verified WB comparison
- PMC8749845 — target-verified WB comparison