WNT7B / Protein Wnt-7b · Western blot design guide

Design a Western Blot for WNT7B

Real validated WNT7B Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-WNT7B 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

Western blot protocol sheet for WNT7B: expected band ~39.3 kDa, hero antibody A05086-2, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **WNT7B** Western blot protocol sheet — expected band ~39.3 kDa, antibody A05086-2, controls and PMC citations. Open the full WNT7B WB guide →

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WNT7B 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	~39.3 kDa
	Observed band	~39 kDa
	Gel	5–20% (catalog A05086-2)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Glycosylated + Cleaved
	Caveat	Secreted protein; lysate sampling
	Gene-set association	MSigDB C7 membership
	Isoform	1 isoform(s)

Section 1

Real Curated WNT7B Western Blot Protocols

The A05086-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human HEK293, human HELA (catalog A05086-2)
Gel %	5–20% (catalog A05086-2)
Load	50ug; reducing conditions (catalog A05086-2)
Transfer	a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A05086-2)
Membrane	Nitrocellulose membrane (catalog A05086-2)
Blocking	5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A05086-2)
Primary antibody	A05086-2 · 0.5 μg/mL (catalog A05086-2)
Primary incubation	overnight at 4°C (catalog A05086-2)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A05086-2)
Secondary incubation	1.5 hour at RT (catalog A05086-2)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A05086-2)
Detection	ECL (catalog A05086-2)

Section 2

What Is the Expected WNT7B Western Blot Band Size?

WNT7B is predicted at 39.3 kDa and observed near 39 kDa; the contribution of signal-peptide cleavage or N-linked glycosylation to migration is not established.

What am I looking at on my blot?
Band near 39 kDa	Matches the empirical WNT7B band; confirm identity with appropriate controls
Band below the precursor position	Could reflect removal of the 1–24 signal peptide; its migration is not established
Little or no band in whole-cell lysate	WNT7B is secreted into the extracellular space and matrix
Broader or higher band	N-linked glycosylation at Asn83, Asn127, or Asn295 could contribute; a visible shift is not established

💡Expected WNT7B appearanceWNT7B has a predicted precursor mass of 39.3 kDa and an empirical band near 39 kDa; confirm band identity with positive and depletion controls because the effects of processing and glycosylation on migration are unquantified.

How each factor affects band size
UniProt predicted precursor mass	Provides a 39.3 kDa sequence-based reference; the empirical band is near 39 kDa
N-linked glycosylation at Asn83	May alter apparent mass, but a site-specific shift is not established
N-linked glycosylation at Asn127	May alter apparent mass, but a site-specific shift is not established
N-linked glycosylation at Asn295	May alter apparent mass, but a site-specific shift is not established
Signal peptide at residues 1–24	Its removal makes the mature protein smaller than the precursor; the migration difference is not established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Secreted WNT7B may be depleted from whole-cell lysate	Examine conditioned medium or extracellular matrix alongside a positive lysate control
Band higher than expected	N-linked glycosylation may affect migration	Compare treated and untreated samples after enzymatic N-glycan removal and verify band identity
Band lower than expected	Signal-peptide removal may reduce mature protein size	Check antibody epitope coverage and compare with a WNT7B depletion control
Broad smear instead of sharp band	Heterogeneous N-linked glycosylation is possible but unproven	Compare glycan-removal conditions and confirm WNT7B specificity
Weak or no signal	Secreted WNT7B may be scarce in the sampled fraction	Test conditioned medium and include a WNT7B-positive lysate control

Source: UniProt P56706 Human Protein Atlas product datasheet (A05086-2)

Experiment evidence

Western blot images showing WNT7B band

A05086-2 · WNT7B

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for WNT7B Western blot

🧪For positive controls for WNT7B in Western blot, you can use a validated WNT7B-positive sample, although the supplied HPA evidence identifies none.

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 alongside the samples.

⚠️Feasibility: WNT7B is secreted, so conditioned medium may give a stronger signal than whole-cell lysate.

HPA tissue expression evidence for WNT7B

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 WNT7B Western Blot Tips

Deeper troubleshooting and optimisation questions for WNT7B, answered from its protein features.

How should WNT7B 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 a second WNT7B band be an isoform?

Isoforms · The supplied record lists one isoform and no alternative sequence. An additional band therefore cannot be assigned to a documented WNT7B isoform from these features alone.

Which WNT7B glycosylation sites matter when assessing bands?

PTM · UniProt lists N-linked glycosylation at Asn83, Asn127, and Asn295. These are UniProt full-length sequence coordinates; antibody or paper numbering may differ. Consider these sites when investigating band heterogeneity, without assuming they cause a particular band.

Does this guide establish induction of WNT7B?

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 WNT7B?

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 A05086-2 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.

What should be considered when quantifying WNT7B bands?

Quantitation · WNT7B is secreted and can occur in a soluble 1:1 complex with AFM. Compare the same sample fraction and preparation conditions across samples, and report which fraction was measured. A band from one fraction does not by itself quantify all extracellular WNT7B.

Why is WNT7B observed near 39 kDa?

Interpretation · The reported band near 39 kDa is close to the 39.3 kDa predicted mass. WNT7B has a signal peptide at residues 1–24 and three listed N-linked glycosylation sites, but those features alone do not establish a visible shift or explain the apparent mass.

WNT7B is listed as secreted and present in the extracellular space and extracellular matrix. When choosing samples, consider both extracellular material and matrix-associated material; a cellular fraction alone may not capture the same WNT7B pool.

WNT7B has 11 listed disulfide bonds, three N-linked glycosylation sites, and a reported soluble complex with AFM. These features warrant checking sample preparation and band identity, but none alone identifies the cause of an unexpected band.

Boster reagents

WNT7B 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

Western blot analysis of WNT7B using anti-WNT7B antibody (A05086-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HEK293 whole cell lysates, Lane 2: human HELA whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-WNT7B antigen affinity purified polyclonal antibody (Catalog # A05086-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for WNT7B at approximately 39KD. The expected band size for WNT7B is at 39KD.

Anti-WNT7B Antibody Picoband®

Cat # A05086-2

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The catalog reports one anti-WNT7B antibody for Western blotting: A05086-2, reported reactive with human samples. Its product image shows an approximately 39 kDa band in reducing HEK293 and HeLa whole-cell lysates. No independent validation is supplied.

Which to pick: A05086-2 is the only listed option. Its WB image uses human HEK293 and HeLa whole-cell lysates, so it is the documented choice for those tested contexts; performance in other samples is not established here.

Source: BosterBio WNT7B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P56706.
Human Protein Atlas. WNT7B tissue expression.
PMC7829129 — target-verified WB comparison
PMC7893751 — target-verified WB comparison
PMC11529306 — target-verified WB comparison