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- Table of Contents
AXIN2 / Axin-2 · Western blot design guide
Design a Western Blot for AXIN2
Real validated AXIN2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AXIN2 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
AXIN2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~93.6 kDa | |
| Observed band | ~100 kDa | |
| Gel | 5–20% (catalog A01772-2) | |
| Positive control | Breast (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated AXIN2 Western Blot Protocols
The A01772-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human PC-3, human MDA-MB-453 (catalog A01772-2) |
| Gel % | 5–20% (catalog A01772-2) |
| Load | 30 ug; reducing conditions (catalog A01772-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01772-2) |
| Membrane | nitrocellulose membrane (catalog A01772-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01772-2) |
| Primary antibody | A01772-2 · 0.5 μg/mL (catalog A01772-2) |
| Primary incubation | overnight at 4°C (catalog A01772-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01772-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01772-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01772-2) |
| Detection | ECL (catalog A01772-2) |
Section 2
What Is the Expected AXIN2 Western Blot Band Size?
AXIN2 is predicted at 93.6 kDa and observed near 100 kDa; the cause of the difference is not established.
| Band near 100 kDa | Matches the reported AXIN2 band in whole-cell lysate |
| Band near 93.6 kDa | Near the sequence-predicted AXIN2 mass; confirm its identity |
| Band in whole-cell lysate | Consistent with cytoplasmic AXIN2 |
| Additional band below 100 kDa | Its identity is unestablished by the supplied features |
💡Expected AXIN2 appearanceAXIN2 has a predicted mass of 93.6 kDa and a reported band near 100 kDa in reducing whole-cell lysates; the difference has no established cause here, so confirm band identity with ordinary controls.
| UniProt predicted mass | Provides a 93.6 kDa sequence-based reference |
| Predicted molecular weight in daltons | Gives the same mass reference as 93,568 Da |
| Predicted mass versus reported migration | The reported band near 100 kDa runs above the 93.6 kDa prediction |
| Predicted mass as an identity reference | A band near 93.6 kDa still requires identity confirmation |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | AXIN2 may be below detection in the sampled lysate | Check loading and use a positive-control lysate |
| Band higher than expected | The cause of migration above 93.6 kDa is unestablished | Compare with the reported 100 kDa band and confirm identity by AXIN2 knockdown |
| Band lower than expected | Identity of the lower band is uncertain | Confirm identity by AXIN2 knockdown |
| Multiple bands | Additional band identities are unestablished | Compare with a positive control and AXIN2 knockdown |
| Weak or no signal | AXIN2 signal may be below the assay detection limit | Check loading, transfer, and antibody performance with a positive control |
Experiment evidence
Western blot images showing AXIN2 band
A01772-2 · AXIN2
M01772 · AXIN2
A01772 · AXIN2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AXIN2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AXIN2 in Western blot, you can use breast tissue, which HPA scores High.
Positive control: Breast (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: No tissue is listed as Not detected, so use siRNA knockdown or a KO line for a clean negative control.
HPA tissue expression evidence for AXIN2
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 |
|---|---|---|---|---|
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Prostate | glandular cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced AXIN2 Western Blot Tips
Deeper troubleshooting and optimisation questions for AXIN2, answered from its protein features.
How should AXIN2 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 AXIN2 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning additional bands to specific AXIN2 isoforms.
Which modifications should I consider when interpreting AXIN2 bands?
PTM · The record lists phosphoprotein, ADP-ribosylation, and ubiquitin-like conjugation keywords, without residue-specific modification entries. Treat these as possible considerations, not proof that a particular band represents modified AXIN2.
Should Wnt stimulation increase the AXIN2 band?
Induction · AXIN2 is annotated with the Wnt signaling pathway, but the supplied features do not establish how its abundance responds to stimulation. Compare stimulated and control samples before attributing a band change to Wnt signaling.
What transfer method to use for AXIN2 Western blot?
Transfer · Use a transfer setup that efficiently recovers a protein appearing near 100 kDa, then check transfer at that size. The supplied features do not identify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01772-2 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 I quantify the AXIN2 band?
Quantitation · Quantify the approximately 100 kDa band consistently across samples, using exposures within a measurable range. Because the record also lists protein modifications, avoid combining additional bands with the main band unless their AXIN2 identity is established.
Why might AXIN2 appear near 100 kDa instead of 93.6 kDa?
Interpretation · The supplied apparent band is approximately 100 kDa, while the predicted mass is 93.6 kDa. AXIN2 is annotated as a phosphoprotein, ADP-ribosylated protein, and ubiquitin-like conjugation target, but these annotations alone do not explain the difference or establish a visible shift.
Compare them with the approximately 100 kDa observed band. The single listed isoform does not explain multiple bands, and modification keywords do not establish the identity of any extra band. Verify band identity before interpreting changes.
Compare them with the approximately 100 kDa observed band. The single listed isoform does not explain multiple bands, and modification keywords do not establish the identity of any extra band. Verify band identity before interpreting changes.
Boster reagents
AXIN2 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-Axin 2/AXIN2 Antibody Picoband®
Cat # A01772-2
Real WB data
Anti-Axin2/Conductin Rabbit Monoclonal Antibody
Cat # M01772
Real WB data
Anti-Axin-2 AXIN2 Antibody
Cat # A01772
Three the supplier anti-AXIN2 antibodies have Western blot images. The captions document specific cell lysates or mouse lung, with detailed conditions for A01772-2. These examples do not establish performance across every listed species or sample type.
Which to pick: For human cell lysates, A01772-2 shows four human lines with detailed conditions. A01772 shows mouse lung. M01772 lists human, mouse, and rat reactivity and shows SW480 and PC-12 lysates; choose according to your sample and verify performance in your assay.
Source: BosterBio AXIN2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y2T1.
- Human Protein Atlas. AXIN2 tissue expression.
- PMC13591777 — target-verified WB comparison
- PMC12630608 — target-verified WB comparison
- PMC3950968 — target-verified WB comparison