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- Table of Contents
WNT10B / Protein Wnt-10b · Western blot design guide
Design a Western Blot for WNT10B
Real validated WNT10B Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-WNT10B 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
WNT10B 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 | ~43 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Stomach (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 | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated WNT10B Western Blot Protocols
The A02574 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human skeletal muscle tissue lysate (catalog A02574) |
| 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 | A02574 · (A) 2 and (B) 4 μg/mL (catalog A02574) |
| 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 WNT10B Western Blot Band Size?
WNT10B has a predicted 43 kDa precursor; secretion, signal-peptide cleavage, glycosylation, and isoforms could affect its blot, but their migration effects are unverified.
| Band near 43 kDa | consistent with the predicted WNT10B precursor mass; identity requires confirmation |
| Band above 43 kDa | may reflect N-linked glycosylation at Asn93 or Asn335; migration effect is unverified |
| Band below 43 kDa | may reflect removal of the 1–28 signal peptide; mature mass is unspecified |
| Several bands | could include isoforms 1 and 2; their migration difference is unknown |
| Little or no band in lysate | consistent with secretion into the extracellular space or matrix |
💡Expected WNT10B appearanceUniProt predicts a 43 kDa WNT10B precursor; signal-peptide removal, N-linked glycosylation, and isoforms could affect migration, but no empirical band size is supplied, so confirm band identity with appropriate controls.
| UniProt predicted mass | 43 kDa for the precursor; apparent migration is unverified |
| N-linked glycosylation at Asn93 | may increase apparent size if the site is occupied |
| N-linked glycosylation at Asn335 | may increase apparent size if the site is occupied |
| Signal peptide at residues 1–28 | cleavage produces a smaller mature protein than the precursor |
| Splice isoforms 1 and 2 | may differ in size; relative masses and separation are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | WNT10B is secreted into the extracellular space and matrix | check conditioned medium or extracellular matrix fractions alongside lysate |
| Band higher than expected | N-linked glycosylation at Asn93 or Asn335 may alter migration | compare treated and untreated samples after N-glycan removal and verify band identity |
| Band lower than expected | cleavage of the 1–28 signal peptide may reduce size | compare precursor and secreted fractions and verify the band with another antibody |
| Broad smear instead of sharp band | variable N-linked glycosylation is possible but unconfirmed | compare treated and untreated samples after N-glycan removal |
| Multiple bands | isoforms 1 and 2 exist, but their migration pattern is unknown | use isoform controls or an independent antibody to identify the bands |
| Weak or no signal | secretion may leave little WNT10B in the sampled lysate | test conditioned medium and a positive control |
Experiment evidence
Western blot images showing WNT10B band
A02574 · WNT10B
A02574-1 · WNT10B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for WNT10B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for WNT10B in Western blot, you can use stomach tissue, which shows high expression in HPA.
Positive control: Stomach (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: WNT10B is secreted, so whole-cell lysates may give weak signal; consider conditioned medium.
HPA tissue expression evidence for WNT10B
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 |
|---|---|---|---|---|
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | 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 WNT10B Western Blot Tips
Deeper troubleshooting and optimisation questions for WNT10B, answered from its protein features.
How should WNT10B 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.
How could WNT10B isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, residues 172–191 are replaced and residues 192–389 are missing, so isoform 2 may produce a smaller band. Check whether the antibody recognizes a region retained in the isoform being measured. Coordinates here follow the supplied UniProt canonical sequence.
Which glycosylation sites matter when comparing WNT10B bands?
PTM · The supplied UniProt features list N-linked sites at Asn93 and Asn335 in the canonical sequence. Asn93 lies in a region retained in isoform 2; Asn335 lies in the region missing from isoform 2. These annotations can guide interpretation, but they do not establish the size of any band shift.
Yes. UniProt lists phosphothreonine at canonical residue 46, a position retained in isoform 2. Use that coordinate when checking antibody documentation, since numbering conventions may differ. The annotation alone does not show that phosphorylation changes WNT10B migration or abundance in your sample.
Yes. UniProt lists phosphothreonine at canonical residue 46, a position retained in isoform 2. Use that coordinate when checking antibody documentation, since numbering conventions may differ. The annotation alone does not show that phosphorylation changes WNT10B migration or abundance in your sample.
Does this guide establish induction of WNT10B?
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.
How should transfer be checked for WNT10B?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02574 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.
What should WNT10B quantitation account for?
Quantitation · Compare the same sample compartment and preparation across conditions: UniProt describes WNT10B as secreted and extracellular matrix associated. Its two isoforms differ substantially in sequence, so confirm that the antibody recognizes the form being quantified. Record which band or bands contribute to the measurement.
Why might WNT10B migrate differently from its predicted 43 kDa mass?
Interpretation · The 43 kDa value is predicted for the precursor. UniProt lists a signal peptide at residues 1–28 and N-linked glycosylation sites at Asn93 and Asn335. Processing and glycosylation could affect apparent mass, but these features alone do not establish a visible shift. No observed band size is supplied.
UniProt places WNT10B in the extracellular space and extracellular matrix and identifies it as secreted. Consider those compartments when selecting samples and interpreting a weak cellular signal. The listed 1:1 soluble complex with AFM is also relevant to interpreting extracellular WNT10B.
First compare the bands with the documented isoforms and the antibody’s recognition region. Also consider the 1–28 signal peptide, Asn93 and Asn335 glycosylation sites, and 11 listed disulfide bonds when reviewing sample preparation. These features suggest possibilities but do not identify an unexpected band without additional evidence.
UniProt places WNT10B in the extracellular space and extracellular matrix and identifies it as secreted. Consider those compartments when selecting samples and interpreting a weak cellular signal. The listed 1:1 soluble complex with AFM is also relevant to interpreting extracellular WNT10B.
First compare the bands with the documented isoforms and the antibody’s recognition region. Also consider the 1–28 signal peptide, Asn93 and Asn335 glycosylation sites, and 11 listed disulfide bonds when reviewing sample preparation. These features suggest possibilities but do not identify an unexpected band without additional evidence.
Boster reagents
WNT10B 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 Wnt-10b Wnt10b Antibody
Cat # A02574
Real WB data
Anti-WNT10B Antibody Picoband®
Cat # A02574-1
Both listed anti-WNT10B antibodies have Western blot images from human samples and list Human, Mouse, and Rat reactivity. The supplied images do not establish performance in mouse or rat samples. For A02574-1, the reported band is approximately 50 kDa versus an expected 43 kDa.
Which to pick: Choose A02574 for a human skeletal muscle example tested at 2 or 4 μg/mL. Choose A02574-1 for human T-47D and HeLa whole cell lysate examples tested at 0.5 μg/mL. Both have WB images; select based on the sample closest to yours.
Source: BosterBio WNT10B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00744.
- Human Protein Atlas. WNT10B tissue expression.
- PMC7859977 — target-verified WB comparison
- PMC5452953 — target-verified WB comparison
- PMC5572676 — target-verified WB comparison