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- Table of Contents
FOXF1 / Forkhead box protein F1 · Western blot design guide
Design a Western Blot for FOXF1
Real validated FOXF1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FOXF1 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
FOXF1 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 | ~40.1 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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated FOXF1 Western Blot Protocols
The M03563 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog M03563) |
| 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 | M03563; 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 FOXF1 Western Blot Band Size?
FOXF1 is predicted at 40.1 kDa; no empirical band is supplied, and the listed features do not demonstrate altered migration.
| Single band near 40.1 kDa | consistent with the UniProt predicted mass; confirm identity with controls |
| Band enriched in a nuclear fraction | consistent with FOXF1 nuclear localization |
| Weak band in whole-cell lysate | a nuclear target may be easier to detect after nuclear enrichment |
| Band far from 40.1 kDa | its identity or altered migration needs independent verification |
💡Expected FOXF1 appearanceUniProt predicts FOXF1 at 40.1 kDa, with no empirical band size supplied; a band near that size is plausible, but confirm its identity with appropriate controls.
| UniProt predicted molecular mass | sets a 40.1 kDa reference for the expected band |
| 379-residue sequence | underlies the predicted mass; no separate processed mass is supplied |
| Nuclear localization | may affect detection across fractions, with no established effect on band size |
| Single listed isoform | provides no evidence for distinct isoform band sizes |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | FOXF1 may be difficult to detect in whole-cell lysate | check a nuclear fraction and a suitable positive control |
| Band higher than expected | the identity or migration of a band above 40.1 kDa is unestablished | check antibody specificity and compare with a positive control |
| Band lower than expected | a smaller band is not explained by a listed cleavage feature | check sample integrity and verify band identity |
| Multiple bands | the supplied isoform data do not establish multiple FOXF1 bands | compare controls and test antibody specificity |
| Weak or no signal | limited target abundance or extraction may reduce detection | check loading, transfer, and nuclear enrichment |
| Fragments below expected size | sample degradation is possible; no fragment is documented here | check sample handling and confirm fragments with an independent antibody |
Experiment evidence
Western blot images showing FOXF1 band
M03563 · FOXF1
A03563-1 · FOXF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FOXF1 Western blot
🧪For positive controls for FOXF1 in Western blot, you can use a separately validated FOXF1-positive sample, since HPA provides no positive tissue or cell line.
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 FOXF1.
⚠️Feasibility: HPA has no expression data here, so tissue controls cannot be selected from the supplied evidence.
HPA tissue expression evidence for FOXF1
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 FOXF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for FOXF1, answered from its protein features.
Where should the FOXF1 band appear relative to its predicted mass?
Band shift · FOXF1 has a predicted mass of 40.1 kDa. No observed band position is supplied, so the features cannot establish an expected shift from that value.
Could FOXF1 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to FOXF1 isoforms.
Do listed FOXF1 modifications predict a band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not support a specific modification-based shift; they also cannot rule out modifications not recorded here.
Does this guide establish induction of FOXF1?
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 FOXF1?
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 M03563 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 FOXF1 Western blot signals be quantified?
Quantitation · FOXF1 is nuclear, so use comparable nuclear samples and a consistent loading reference when comparing signals. The supplied features do not establish how abundance changes between conditions.
How should FOXF1 localization guide sample preparation?
Interpretation · FOXF1 is annotated as nuclear. A nuclear-enriched sample may help detect it; compare samples prepared with the same fractionation approach.
Start with the 40.1 kDa predicted mass, then check band specificity experimentally. The supplied record lists one isoform, no alternative sequence, and no modified residues or glycosylation sites, so it cannot identify the cause of an extra band.
Start with the 40.1 kDa predicted mass, then check band specificity experimentally. The supplied record lists one isoform, no alternative sequence, and no modified residues or glycosylation sites, so it cannot identify the cause of an extra band.
Boster reagents
FOXF1 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-FOXF1 Rabbit Monoclonal Antibody
Cat # M03563
Real WB data
Anti-FOXF1 Picoband® Antibody
Cat # A03563-1
Both listed anti-FOXF1 antibodies have Western blot images. M03563 shows HeLa lysate; A03563-1 shows human cell and mouse liver and cell lysates, with a reported 38–40 kDa band. Rat reactivity is listed for A03563-1 but is not shown in its image.
Which to pick: For human HeLa lysate, either has an image. Choose A03563-1 for the shown mouse samples or its broader listed reactivity; choose M03563 if you want the human-reactive monoclonal. Rat is listed only for A03563-1, without a rat WB example.
Source: BosterBio FOXF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q12946.
- Human Protein Atlas. FOXF1 tissue expression.
- PMC7115358 — target-verified WB comparison
- PMC9040237 — target-verified WB comparison
- PMC5243941 — target-verified WB comparison