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- Table of Contents
WNT9A / Protein Wnt-9a · Western blot design guide
Design a Western Blot for WNT9A
Source-linked WNT9A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-WNT9A 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
WNT9A 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 | ~40.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked WNT9A Western Blot Protocol Options
The A07111 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of mouse lung, (catalog A07111) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A07111) |
| Primary antibody | A07111 · 1:1000 (catalog A07111) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A07111) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A07111) |
Section 2
What Is the Expected WNT9A Western Blot Band Size?
WNT9A has a predicted full-length mass of 40.3 kDa; signal-peptide cleavage and Asn103 glycosylation could affect migration, but no empirical band size is supplied.
| Band near 40.3 kDa | consistent with the predicted full-length WNT9A precursor, pending identity controls |
| Band below 40.3 kDa | may reflect cleavage of the 1–29 signal peptide; identity requires confirmation |
| Band migrating above the predicted size | could reflect N-linked glycosylation at Asn103, but its effect on migration is unestablished |
| Little or no band in whole-cell lysate | consistent with secretion of WNT9A into extracellular material |
💡Expected WNT9A appearanceThe predicted full-length WNT9A mass is 40.3 kDa; no empirical band size is supplied, and signal-peptide cleavage or Asn103 glycosylation may alter migration, so confirm band identity with appropriate controls.
| Predicted full-length mass | sets a 40.3 kDa precursor reference, not a measured band position |
| Signal peptide at residues 1–29 | cleavage would make mature WNT9A smaller than the full-length precursor |
| N-linked glycosylation site at Asn103 | glycosylation may increase apparent size; the shift is not established |
| Glycan occupancy at Asn103 | could affect migration if the site is occupied, but distinct bands are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | WNT9A is secreted into extracellular space and matrix | check conditioned medium or extracellular material alongside lysate |
| Band higher than expected | Asn103 glycosylation could affect migration; the cause is unconfirmed | compare with N-glycan removal and verify band identity |
| Band lower than expected | cleavage of the 1–29 signal peptide could lower mature-protein mass | compare precursor and secreted fractions and verify band identity |
| Multiple bands | precursor and signal-peptide-cleaved protein may differ; distinct bands are unproven | compare lysate with conditioned medium and use band-identity controls |
| Weak or no signal | secretion may leave little WNT9A in the sampled lysate | check conditioned medium or extracellular material |
Experiment evidence
Western blot images showing WNT9A band
A07111 · WNT9A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for WNT9A Western blot
🧪For positive controls for WNT9A in Western blot, you can use no HPA-supported tissue or cell line from the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: WNT9A is secreted, so conditioned medium may give a stronger signal than whole-cell lysate.
HPA tissue expression evidence for WNT9A
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced WNT9A Western Blot Tips
Deeper troubleshooting and optimisation questions for WNT9A, answered from its protein features.
How should WNT9A 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 WNT9A isoforms explain multiple bands?
Isoforms · The supplied features list one isoform and no alternative sequence. They provide no specific isoform explanation for multiple bands. Assess processing, glycosylation, and sample fraction before assigning bands to isoforms.
Which WNT9A glycosylation site should I consider?
PTM · UniProt lists one N-linked glycosylation site at Asn-103. This is the coordinate in the supplied full-length sequence; numbering based on the processed protein may differ. Compare untreated and deglycosylated samples if testing whether glycosylation affects a band.
Does this guide establish induction of WNT9A?
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 WNT9A?
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 A07111 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 WNT9A be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might WNT9A migrate differently from its predicted 40.3 kDa?
Interpretation · The 40.3 kDa prediction includes the 1–29 signal peptide. WNT9A also has an N-linked glycosylation site at Asn-103. These features may affect apparent mass, but no observed band size is supplied, so neither establishes a specific shift.
UniProt lists 11 disulfide bonds. Compare matched reducing and nonreducing samples when investigating different migration patterns. A change would support sensitivity to sample conditions, but the feature count alone does not predict a band position.
WNT9A is annotated as secreted and present in extracellular space and extracellular matrix. Examine appropriate extracellular fractions alongside cell-associated material; a weak cell-lysate signal alone does not establish low overall WNT9A abundance.
Keep the sampled compartment and collection conditions consistent. WNT9A is secreted and associated with extracellular space and matrix, so differences between lysate and extracellular fractions can reflect distribution as well as abundance.
UniProt reports a soluble 1:1 WNT9A–AFM complex that prevents oligomerization. Consider this association when interpreting bands under conditions that preserve complexes. The annotation does not establish that a particular Western-blot band contains AFM or predict its apparent mass.
UniProt lists 11 disulfide bonds. Compare matched reducing and nonreducing samples when investigating different migration patterns. A change would support sensitivity to sample conditions, but the feature count alone does not predict a band position.
WNT9A is annotated as secreted and present in extracellular space and extracellular matrix. Examine appropriate extracellular fractions alongside cell-associated material; a weak cell-lysate signal alone does not establish low overall WNT9A abundance.
Keep the sampled compartment and collection conditions consistent. WNT9A is secreted and associated with extracellular space and matrix, so differences between lysate and extracellular fractions can reflect distribution as well as abundance.
UniProt reports a soluble 1:1 WNT9A–AFM complex that prevents oligomerization. Consider this association when interpreting bands under conditions that preserve complexes. The annotation does not establish that a particular Western-blot band contains AFM or predict its apparent mass.
Boster reagents
WNT9A 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-9a WNT9A Antibody
Cat # A07111
The catalog reports A07111, an anti-WNT9A antibody with reported human, mouse, and rat reactivity. Its Western blot image uses mouse lung extract at 1:1000 dilution, with 25 µg per lane. The supplied evidence shows this tested context only.
Which to pick: A07111 is the only listed option and has a Western blot image from mouse lung extract. Its listed reactivity includes human, mouse, and rat; the supplied image documents mouse lung only.
Source: BosterBio WNT9A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O14904.
- Human Protein Atlas. WNT9A tissue expression.
- PMC7814487 — target-verified WB comparison
- PMC5852562 — target-verified WB comparison
- PMC6052404 — target-verified WB comparison