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- Table of Contents
ADORA1 / Adenosine receptor A1 · Western blot design guide
Design a Western Blot for ADORA1
Source-linked ADORA1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ADORA1 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
ADORA1 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 | ~36.5 kDa | |
| Observed band | ~36 kDa | |
| Gel | 10% (catalog M01101) | |
| Positive control | Hippocampus (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked ADORA1 Western Blot Protocol Options
The M01101 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Caco-2, human U2OS, human MCF-7, rat brain, rat C6, mouse brain (catalog M01101) |
| Gel % | 10% (catalog M01101) |
| Load | 30 ug; reducing conditions (catalog M01101) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01101) |
| Membrane | nitrocellulose membrane (catalog M01101) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01101) |
| Primary antibody | M01101 · 1:500 (catalog M01101) |
| Primary incubation | overnight at 4°C (catalog M01101) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M01101) |
| Secondary incubation | 1.5 hour at RT (catalog M01101) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01101) |
| Detection | ECL (catalog M01101) |
Section 2
What Is the Expected ADORA1 Western Blot Band Size?
ADORA1 is predicted at 36.5 kDa and observed at ~36 kDa; the cause of the small difference is not established.
| Band at ~36 kDa | Matches the empirical ADORA1 band; confirm identity with controls |
| Band above ~36 kDa | May reflect modification at N-linked glycosylation site Asn159; the site alone does not establish a visible shift |
| Several bands | Isoforms 1 and 2 are annotated, but distinct band positions are unconfirmed |
| Faint or absent band in lysate | ADORA1 is a multi-pass membrane protein that may be poorly extracted |
💡Expected ADORA1 appearanceUniProt predicts 36.5 kDa, while antibody QC shows a band at ~36 kDa; confirm band identity with appropriate controls because the supplied features do not establish its migration mechanism.
| UniProt predicted mass | 36.5 kDa provides the sequence-based reference; the observed band is ~36 kDa |
| N-linked glycosylation site Asn159 | Modification could alter apparent size, but a visible shift is not established |
| Splice isoform 1 | Its individual mass and migration are not supplied |
| Splice isoform 2 | Its size relative to isoform 1 and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ADORA1 may be poorly extracted | Check membrane extraction and use an ADORA1-positive lysate |
| Band higher than expected | Asn159 glycosylation is possible, but the band identity is unproven | Compare treated and untreated lysates and confirm identity by ADORA1 depletion |
| Band lower than expected | Isoform identity or another cause is unresolved | Confirm the band by ADORA1 depletion and assess isoform expression |
| Multiple bands | Isoforms 1 and 2 exist, but their band positions are unknown | Use ADORA1 depletion and isoform-specific controls to assign bands |
| Weak or no signal | Extraction of multi-pass membrane ADORA1 may be inefficient | Check the membrane fraction and include a positive control |
Experiment evidence
Western blot images showing ADORA1 band
M01101 · ADORA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ADORA1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ADORA1 in Western blot, you can use hippocampus tissue, which HPA scores High.
Positive control: Hippocampus (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, ADORA1 may require membrane-enriched lysate for a clear signal.
HPA tissue expression evidence for ADORA1
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 |
|---|---|---|---|---|
| Hippocampus | neuronal projections | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADORA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ADORA1, answered from its protein features.
How should ADORA1 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could ADORA1 isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Relative to the canonical sequence, isoform 2 replaces residues 115..125 and lacks residues 126..326, so it could migrate differently if expressed. Check whether the antibody epitope is retained before assigning a band to isoform 2.
How should ADORA1 glycosylation be assessed?
PTM · The listed N-linked site is Asn159 in UniProt canonical numbering. Isoform 2 lacks that position. Compare appropriately treated and untreated samples if testing glycosylation, but do not assign a band shift from the site annotation alone. Antibody or paper numbering may differ from UniProt numbering.
Does this guide establish induction of ADORA1?
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 ADORA1 Western blot?
Transfer · ADORA1 is a multi-pass cell-membrane protein. Check recovery of the approximately 36 kDa region on the membrane and in the post-transfer gel, then adjust transfer conditions if needed. The supplied features do not identify one required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01101 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 can ADORA1 bands be quantified consistently?
Quantitation · ADORA1 is annotated as a cell-membrane protein, so use comparable sample preparation and loading across samples. Quantify the same defined band or bands each time, and record whether the antibody can detect both annotated isoforms.
Why is the ADORA1 band near its predicted mass?
Interpretation · The observed band is about 36 kDa, close to the 36.5 kDa predicted mass. UniProt lists N-linked glycosylation at Asn159, but that feature alone does not establish a visible shift or explain a mass difference.
Compare them with the approximately 36 kDa observed band. Isoform 2 lacks canonical residues 126..326, and canonical Asn159 is annotated as glycosylated; these are candidates to investigate, not band assignments. Check antibody epitope coverage and sample preparation before identifying an unexpected band.
Compare them with the approximately 36 kDa observed band. Isoform 2 lacks canonical residues 126..326, and canonical Asn159 is annotated as glycosylated; these are candidates to investigate, not band assignments. Check antibody epitope coverage and sample preparation before identifying an unexpected band.
Boster reagents
ADORA1 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-ADORA1 Rabbit Monoclonal Antibody
Cat # M01101
M01101 is a rabbit monoclonal anti-ADORA1 antibody with a Western blot image showing an approximately 36 kDa band. The caption reports human cell, rat cell and brain, and mouse brain lysates; catalogued reactivity is Human.
Which to pick: M01101 is the only listed option. It has a Western blot image and catalogued Human reactivity. The image includes rat and mouse samples, but those examples do not establish general reactivity across either species.
Source: BosterBio ADORA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P30542.
- Human Protein Atlas. ADORA1 tissue expression.
- PMC7520255 — target-verified WB comparison
- PMC6471230 — target-verified WB comparison
- PMC5225479 — target-verified WB comparison