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- Table of Contents
CNR2 / Cannabinoid receptor 2 · Western blot design guide
Design a Western Blot for CNR2
Source-linked CNR2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CNR2 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
CNR2 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 | ~39.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked CNR2 Western Blot Protocol Options
The A01307-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01307-3; use the WB datasheet starting dilution (standard starting point) |
| 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 CNR2 Western Blot Band Size?
CNR2 is predicted at 39.7 kDa; Asn11 glycosylation could affect migration, but no empirical band size or visible shift is established.
| Band near 39.7 kDa | Consistent with the predicted CNR2 mass; confirm identity with controls. |
| Band above 39.7 kDa | N-linked glycosylation at Asn11 could affect migration, but a visible shift is not established. |
| Weak band in a soluble fraction | CNR2 is a multi-pass membrane protein and may be poorly recovered in this fraction. |
| Single sharp band | Consistent with one annotated isoform; Asn11 glycosylation need not produce a visible second band. |
💡Expected CNR2 appearanceCNR2 has a predicted mass of 39.7 kDa, but no empirical band size is supplied; confirm any candidate band with appropriate identity controls.
| Predicted molecular mass | Places the 360-residue CNR2 sequence at 39.7 kDa before any possible migration effects. |
| Asn11 N-linked glycosylation site | Could increase apparent size if occupied; no shift is demonstrated. |
| Asn11 glycan occupancy | Could differ between molecules, but distinct bands are not established. |
| Full-length 360-residue sequence | Provides the basis for the predicted 39.7 kDa mass; no cleavage is annotated. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The multi-pass membrane protein may be poorly recovered. | Check the membrane fraction and extraction conditions. |
| Band higher than expected | Asn11 N-linked glycosylation could affect migration. | Compare treated and untreated samples with a CNR2 identity control. |
| Band lower than expected | Its identity is uncertain; no signal peptide or propeptide cleavage is annotated. | Check specificity with CNR2 depletion and a size marker. |
| Multiple bands | Their identities are uncertain; one isoform is annotated. | Compare with CNR2 depletion and assess the Asn11 glycosylation contribution. |
| Weak or no signal | Membrane extraction or low CNR2 recovery may limit detection. | Check membrane enrichment, loading, and antibody performance. |
| Fragments below expected size | Sample degradation is possible; no specific fragments are annotated. | Use fresh samples and protease inhibitors, then verify CNR2 identity. |
Experiment evidence
Western blot images showing CNR2 band
A01307-3 · CNR2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CNR2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CNR2 in Western blot, you can use appendix tissue, which HPA scores as highly expressed.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, CNR2 may be easier to detect in membrane-enriched samples.
HPA tissue expression evidence for CNR2
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced CNR2 Western Blot Tips
Deeper troubleshooting and optimisation questions for CNR2, answered from its protein features.
How should CNR2 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 multiple CNR2 bands represent annotated isoforms?
Isoforms · UniProt lists one CNR2 isoform and no alternative sequence in the supplied record. Do not assign multiple bands to annotated isoforms on this evidence alone; assess their identity with appropriate controls.
How can glycosylation affect CNR2 band interpretation?
PTM · UniProt annotates one N-linked glycosylation site at Asn11. If bands differ in apparent mass, compare matched samples before and after deglycosylation and assess whether the pattern changes. The annotation alone does not establish that a particular band is glycosylated.
The supplied UniProt features list phosphoserine at positions 335, 336, and 352, and phosphothreonine at 338. Match a phospho-specific antibody to the intended site and verify its numbering convention; UniProt coordinates may differ from paper or antibody numbering. These annotations do not predict a visible band shift.
The supplied UniProt features list phosphoserine at positions 335, 336, and 352, and phosphothreonine at 338. Match a phospho-specific antibody to the intended site and verify its numbering convention; UniProt coordinates may differ from paper or antibody numbering. These annotations do not predict a visible band shift.
Does this guide establish induction of CNR2?
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 CNR2 Western blot?
Transfer · CNR2 is a 360-residue, multi-pass membrane protein with a predicted mass of 39.7 kDa. Compare transfer conditions on matched samples and verify transfer with a total-protein stain. The supplied features do not identify a preferred transfer method.
How should blocking be optimized for CNR2 detection?
Blocking · Compare blockers on matched samples and choose the condition with clear CNR2 signal and low background. If detecting one of the annotated phosphorylation sites, evaluate the blocker with that phospho-specific antibody as well as the total-CNR2 antibody; their performance may differ.
How should CNR2 bands be quantified?
Quantitation · Quantify a validated CNR2 band within the assay's linear signal range and normalize across matched samples. For a phosphorylation measurement, compare site-specific signal with total CNR2 signal. The supplied features identify sites but do not establish how strongly each is modified in a sample.
Should CNR2 migrate at its predicted 39.7 kDa?
Interpretation · Use 39.7 kDa as the predicted mass, not a guaranteed band position. CNR2 has an annotated N-linked glycosylation site at UniProt Asn11 and four phosphorylation sites, but those features alone do not establish a visible shift or explain any observed mass difference.
Check band identity with suitable controls before assigning a cause. The supplied record lists N-linked glycosylation at UniProt Asn11 and phosphorylation at 335, 336, 338, and 352, but neither feature proves an unexpected band results from modification. It lists no alternative sequence to support an annotated isoform assignment.
Check band identity with suitable controls before assigning a cause. The supplied record lists N-linked glycosylation at UniProt Asn11 and phosphorylation at 335, 336, 338, and 352, but neither feature proves an unexpected band results from modification. It lists no alternative sequence to support an annotated isoform assignment.
Boster reagents
CNR2 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-CB2 Antibody (C-term)
Cat # A01307-3
A01307-3 is an anti-CNR2 (CB2) C-terminal antibody listed for human reactivity. Its WB image shows detection in A431 cell line lysate at 35 µg per lane. The supplied evidence does not establish performance in other samples.
Which to pick: A01307-3 is the only listed option. Choose it for a human CNR2 blot when the A431 lysate WB image provides a relevant starting point; verify performance in your own sample and conditions.
Source: BosterBio CNR2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P34972.
- Human Protein Atlas. CNR2 tissue expression.
- PMC5444694 — target-verified WB comparison
- PMC7552904 — target-verified WB comparison
- PMC10725463 — target-verified WB comparison