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- Table of Contents
FZD7 / Frizzled-7 · Western blot design guide
Design a Western Blot for FZD7
Real validated FZD7 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FZD7 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
FZD7 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 | ~63.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 + Ubl conjugation | |
| Caveat | Peptide-blocking control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated FZD7 Western Blot Protocols
The A04391 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HELA cells (catalog A04391) |
| 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 | A04391 · 1:2000 (catalog A04391) |
| 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 FZD7 Western Blot Band Size?
FZD7 has a predicted full-length mass of 63.6 kDa; cleavage and N-linked glycosylation could affect migration, but no empirical band size demonstrates either effect.
| Band near 63.6 kDa | Consistent with the predicted full-length FZD7 mass; confirm band identity. |
| Band below 63.6 kDa | Could reflect signal-peptide cleavage; confirm band identity. |
| Band above 63.6 kDa | N-linked glycosylation at Asn63 or Asn164 could affect migration; verify experimentally. |
| Doublet near the predicted size | Could reflect precursor and signal-peptide-cleaved forms; verify both bands. |
💡Expected FZD7 appearanceUniProt predicts a 63.6 kDa full-length precursor; signal-peptide cleavage and N-linked glycosylation could alter migration, but no empirical FZD7 band size is supplied, so confirm identity with appropriate controls.
| Predicted full-length mass | 63.6 kDa is the sequence-based reference, not a measured band. |
| N-linked glycosylation at Asn63 | Could increase apparent size if modified; the shift is not established. |
| N-linked glycosylation at Asn164 | Could increase apparent size if modified; the shift is not established. |
| Signal peptide at residues 1–32 | Cleavage would make mature FZD7 smaller than the full-length precursor; its band size is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | FZD7 is a multi-pass membrane protein and may be poorly recovered. | Check membrane-protein extraction and use a positive-control lysate. |
| Band higher than expected | N-linked glycosylation could affect migration; the extent is unknown. | Compare untreated and deglycosylated samples and verify band identity. |
| Band lower than expected | Signal-peptide cleavage could lower the mass relative to the precursor. | Check whether the antibody recognizes the mature protein and verify band identity. |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unproven. | Compare with a deglycosylated sample and check sample quality. |
| Multiple bands | Precursor and mature forms are possible, but their resolution is unknown. | Use peptide competition or an independent antibody to identify FZD7 bands. |
| Weak or no signal | Membrane extraction may yield too little FZD7. | Check extraction efficiency and antibody performance with a positive control. |
Experiment evidence
Western blot images showing FZD7 band
A04391 · FZD7
A04391 · FZD7
A04391Y87 · FZD7
A04391Y87 · FZD7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FZD7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FZD7 in Western blot, you can use adrenal gland tissue, where HPA reports high expression.
Positive control: Adrenal gland (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: As a multi-pass membrane protein, FZD7 may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for FZD7
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | High | 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 → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced FZD7 Western Blot Tips
Deeper troubleshooting and optimisation questions for FZD7, answered from its protein features.
How should FZD7 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.
Should multiple FZD7 bands be assigned to annotated isoforms?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign additional bands to annotated FZD7 isoforms on this evidence. Check band specificity before interpreting multiple bands.
How can FZD7 glycosylation be assessed on a blot?
PTM · FZD7 has annotated N-linked glycosylation at Asn63 and Asn164 in UniProt numbering. Compare matched samples before and after an N-glycan removal treatment, alongside an untreated control. A mobility change would support glycosylation as a contributor, but the annotations alone do not predict the size of that change.
Does this guide establish induction of FZD7?
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 FZD7 Western blot?
Transfer · FZD7 is a multipass protein found at cell and endosome membranes. Check transfer efficiency for the FZD7 band in your membrane-protein preparation and adjust transfer conditions if protein remains in the gel. The supplied features do not identify one preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04391 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 FZD7 band intensity be quantified?
Quantitation · Compare the same validated FZD7 band across samples prepared and blotted under consistent conditions. Account for loading and transfer when comparing intensities. Its membrane localization and annotated glycosylation sites make consistent preparation especially useful; the features do not identify a specific band position.
Why might FZD7 migrate differently from its predicted 63.6 kDa?
Interpretation · The 63.6 kDa value is calculated from the sequence. FZD7 has a signal peptide at residues 1–32 and annotated N-linked glycosylation sites at Asn63 and Asn164, using UniProt coordinates. These features could affect the protein analyzed, but they do not establish a visible shift or explain any particular band. No empirical band position was supplied.
UniProt annotates a signal peptide at residues 1–32. Account for this region when comparing the full-sequence predicted mass with a processed protein. The feature does not establish where a Western blot band will migrate.
First verify antibody specificity and check transfer and sample preparation. FZD7 has a signal peptide, two N-linked glycosylation sites, and five annotated disulfide bonds, but these features do not identify an unexpected band by themselves. Use matched glycan-removal or reducing-condition comparisons where relevant, and report any assignment as tentative until validated.
UniProt annotates a signal peptide at residues 1–32. Account for this region when comparing the full-sequence predicted mass with a processed protein. The feature does not establish where a Western blot band will migrate.
First verify antibody specificity and check transfer and sample preparation. FZD7 has a signal peptide, two N-linked glycosylation sites, and five annotated disulfide bonds, but these features do not identify an unexpected band by themselves. Use matched glycan-removal or reducing-condition comparisons where relevant, and report any assignment as tentative until validated.
Boster reagents
FZD7 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-7 FZD7 Antibody
Cat # A04391
Real WB data
Anti-Frizzled-7 (Y87) FZD7 Antibody
Cat # A04391Y87
Two anti-FZD7 antibodies have WB images: A04391 with HeLa cells at 1:2000, and A04391Y87 with HeLa, BV2, and PC12 whole-cell lysates at 1:500. These examples show tested contexts, not comprehensive validation across the listed species.
Which to pick: For a rat sample, consider A04391Y87, the only antibody listed with rat reactivity; its WB image includes PC12 lysate. For human or mouse samples, compare the reported conditions: A04391 shows HeLa at 1:2000, while A04391Y87 shows HeLa, BV2, and PC12 at 1:500.
Source: BosterBio FZD7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75084.
- Human Protein Atlas. FZD7 tissue expression.
- PMC7487719 — target-verified WB comparison
- PMC13411977 — target-verified WB comparison
- PMC5349891 — target-verified WB comparison