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- Table of Contents
FEV / Protein FEV · Western blot design guide
Design a Western Blot for FEV
Source-linked FEV Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FEV 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
FEV 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 | ~25 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
Source-Linked FEV Western Blot Protocol Options
The A03460 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03460; 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 FEV Western Blot Band Size?
FEV is predicted at 25 kDa; no empirical band is supplied, and the listed features do not establish altered migration.
| Single band near 25 kDa | Consistent with FEV's predicted mass; confirm its identity |
| Band near 25 kDa in a nuclear fraction | Consistent with FEV's predicted mass and nuclear location |
| Stronger band near 25 kDa in a nuclear fraction | Consistent with FEV's nuclear location |
| Little signal in a cytoplasmic fraction | Consistent with FEV's nuclear location |
💡Expected FEV appearanceFEV has a predicted mass of 25 kDa, but no empirical band size is supplied; confirm a band near that position with ordinary identity controls.
| UniProt predicted mass | Places full-length FEV near 25 kDa |
| Calculated mass of 25,030 Da | Corresponds to approximately 25 kDa |
| Predicted full-length mass | Provides a reference for assessing apparent migration |
| Predicted 25 kDa band position | Does not establish the experimentally observed position |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear FEV may be poorly recovered | Check nuclear fraction recovery and assay sensitivity |
| Band higher than expected | Identity of the band is unestablished | Compare nuclear fractions and use a FEV depletion control |
| Band lower than expected | Identity of the band is unestablished | Use a FEV depletion control and check sample integrity |
| Multiple bands | The supplied features do not establish distinct FEV band sizes | Use a FEV depletion control to identify the specific band |
| Weak or no signal | Nuclear FEV may be diluted in whole-cell lysate | Test a nuclear-enriched preparation and verify antibody performance |
Experiment evidence
Western blot images showing FEV band
A03460 · FEV
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FEV Western blot
🧪For positive controls for FEV in Western blot, you can use no HPA-supported sample because no positive tissue or cell data were supplied.
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: HPA supplies no tissue data, so tissue-based controls cannot be selected; an FEV knockdown or KO line could provide a negative control.
HPA tissue expression evidence for FEV
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 FEV Western Blot Tips
Deeper troubleshooting and optimisation questions for FEV, answered from its protein features.
Where should the FEV band appear relative to its predicted mass?
Band shift · FEV has a predicted mass of 25 kDa. No observed band position is supplied, so use 25 kDa as a reference rather than an exact expected position. The supplied features do not establish a band shift.
Could FEV isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning additional bands to FEV isoforms; verify their identity before interpreting them.
Do annotated modifications explain a shifted FEV band?
PTM · No modified residues or glycosylation sites are listed for FEV. The supplied features therefore provide no specific modification to test as the cause of a shift.
Does this guide establish induction of FEV?
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 FEV Western blot?
Transfer · FEV is predicted to be 25 kDa. The supplied features do not identify a transfer method; confirm that protein near 25 kDa transfers and remains on the membrane under your chosen conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03460 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 FEV 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.
Which fraction should be checked for FEV?
Interpretation · FEV is annotated in the nucleus. Check a nuclear fraction when assessing its presence, and compare fraction loading consistently across samples.
FEV is annotated as nuclear. For comparisons, use the same sample fraction across lanes and normalize for loading in that fraction. The supplied features provide no expected change in FEV abundance.
Start with the predicted 25 kDa mass, then verify any additional bands independently. The record lists one isoform and no modified residues or glycosylation sites, so it does not identify a specific cause for extra bands.
FEV is annotated as nuclear. For comparisons, use the same sample fraction across lanes and normalize for loading in that fraction. The supplied features provide no expected change in FEV abundance.
Start with the predicted 25 kDa mass, then verify any additional bands independently. The record lists one isoform and no modified residues or glycosylation sites, so it does not identify a specific cause for extra bands.
Boster reagents
FEV 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 FEV FEV Antibody
Cat # A03460
A03460 is listed for human, mouse, and rat FEV. Its Western blot image shows K562 cell lysate tested with primary antibody at 1:1000 overnight at 4°C. The supplied evidence shows this sample and condition only.
Which to pick: A03460 is the only listed FEV antibody. It has a Western blot image from K562 lysate; the supplied evidence does not show mouse or rat samples tested by Western blot.
Source: BosterBio FEV gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99581.
- Human Protein Atlas. FEV tissue expression.
- PMC4711820 — target-verified WB comparison