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- Table of Contents
FZD10 / Frizzled-10 · Western blot design guide
Design a Western Blot for FZD10
Source-linked FZD10 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FZD10 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
FZD10 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 | ~65.3 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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked FZD10 Western Blot Protocol Options
The A07549 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | COS-7 cells (catalog A07549) |
| 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 | A07549; 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 FZD10 Western Blot Band Size?
FZD10 is predicted at 65.3 kDa; signal-peptide cleavage and N-linked glycosylation could affect migration, but no empirical band position is supplied.
| Band near 65.3 kDa | Consistent with the predicted full-length FZD10 mass; identity needs confirmation |
| Band above 65.3 kDa | N-linked glycosylation at Asn48, Asn153, or Asn485 could affect migration |
| Band below the full-length position | Could reflect cleavage of the 1–20 signal peptide |
| Broad band or smear | Could reflect variation in N-linked glycosylation; the sites alone do not establish a smear |
| Weak or absent band in soluble lysate | FZD10 is a multi-pass cell-membrane protein |
💡Expected FZD10 appearanceFZD10 has a predicted full-length mass of 65.3 kDa, but no empirical band size is supplied; signal-peptide cleavage and N-linked glycosylation may alter migration, so confirm band identity with controls.
| Predicted full-length mass | 65.3 kDa is the sequence-based reference, not a measured band position |
| N-linked glycosylation at Asn48 | May increase apparent band size if glycosylated |
| N-linked glycosylation at Asn153 | May increase apparent band size if glycosylated |
| N-linked glycosylation at Asn485 | May increase apparent band size if glycosylated |
| Signal peptide at residues 1–20 | Cleavage may place mature FZD10 below the full-length precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The multi-pass membrane protein may be poorly recovered in the lysate | Check a membrane-enriched fraction and verify extraction |
| Band higher than expected | N-linked glycosylation may affect migration | Compare treated and untreated samples with a band-identity control |
| Band lower than expected | Signal-peptide cleavage may reduce size | Check antibody epitope coverage and confirm band identity |
| Broad smear instead of sharp band | Variation in N-linked glycosylation is possible | Compare treated and untreated samples and confirm specificity |
| Multiple bands | Processing or glycosylation may produce distinct forms | Use a specificity control before assigning band identities |
Experiment evidence
Western blot images showing FZD10 band
A07549 · FZD10
A07549 · FZD10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FZD10 Western blot
🧪For positive controls for FZD10 in Western blot, you can use an independently validated FZD10-positive sample because HPA provides no positive tissue or cell-line candidate.
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: HPA provides no tissue controls, and this multi-pass membrane protein may require membrane-enriched lysate for a clear signal.
HPA tissue expression evidence for FZD10
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 FZD10 Western Blot Tips
Deeper troubleshooting and optimisation questions for FZD10, answered from its protein features.
How should FZD10 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 alternative FZD10 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning multiple bands to annotated isoforms. Compare each band with appropriate specificity controls before assigning it to FZD10.
Which FZD10 glycosylation sites matter when interpreting bands?
PTM · UniProt annotates N-linked glycosylation at Asn48, Asn153 and Asn485. These are UniProt sequence coordinates; paper or antibody numbering may differ. The annotations identify sites to consider, but do not establish how many are occupied in your sample or whether they produce separate bands.
Does this guide establish induction of FZD10?
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 FZD10 Western blot?
Transfer · FZD10 is a multi-pass cell-membrane protein with a predicted mass of 65.3 kDa. Check transfer efficiency for the band region of interest and confirm that protein remains on the membrane. The supplied UniProt features do not establish a specific transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07549 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 FZD10 bands be quantified across samples?
Quantitation · Because FZD10 is annotated as a multi-pass cell-membrane protein, prepare comparable samples and use the same extraction conditions throughout. Quantify signals within the assay’s linear range and normalize consistently. The supplied features do not identify a validated reference band or establish that every detected band is FZD10.
Why might FZD10 migrate differently from 65.3 kDa?
Interpretation · 65.3 kDa is the predicted mass, and no observed band mass is supplied. FZD10 has a signal peptide at residues 1–20 and three annotated N-linked glycosylation sites. These features may affect the mature protein or its migration, but their presence alone cannot establish a visible shift or explain a measured difference.
UniProt lists five disulfide bonds. Keep reducing conditions consistent across samples when comparing band patterns; a change in those conditions could complicate interpretation. The record does not predict a particular band position under either condition.
Check specificity before assigning them to FZD10. Consider the annotated signal peptide at residues 1–20, N-linked sites at Asn48, Asn153 and Asn485, and five disulfide bonds when comparing sample conditions. These features alone cannot identify an unexpected band, and no empirical FZD10 band size was supplied.
UniProt lists five disulfide bonds. Keep reducing conditions consistent across samples when comparing band patterns; a change in those conditions could complicate interpretation. The record does not predict a particular band position under either condition.
Check specificity before assigning them to FZD10. Consider the annotated signal peptide at residues 1–20, N-linked sites at Asn48, Asn153 and Asn485, and five disulfide bonds when comparing sample conditions. These features alone cannot identify an unexpected band, and no empirical FZD10 band size was supplied.
Boster reagents
FZD10 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-Frizzled-10 FZD10 Antibody
Cat # A07549
The catalog reports one anti-FZD10 antibody for Western blot, A07549. Its catalog entry includes a WB image using COS-7 cells; the supplied evidence does not establish performance across other samples.
Which to pick: A07549 is the only listed option. It reports Human, Monkey, and Mouse reactivity, with a WB image from COS-7 cells; check suitability for your sample and conditions.
Source: BosterBio FZD10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9ULW2.
- Human Protein Atlas. FZD10 tissue expression.
- PMC7476091 — target-verified WB comparison