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- Table of Contents
KIF14 / Kinesin-like protein KIF14 · Western blot design guide
Design a Western Blot for KIF14
Source-linked KIF14 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KIF14 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
KIF14 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 | ~186.5 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked KIF14 Western Blot Protocol Options
The A04137 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A04137) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04137) |
| Primary antibody | A04137 · 1:1000 (catalog A04137) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04137) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04137) |
Section 2
What Is the Expected KIF14 Western Blot Band Size?
Full-length KIF14 is predicted at 186.5 kDa; phosphorylation is documented, but its effect on migration has not been demonstrated here.
| Band near 186.5 kDa | Consistent with full-length KIF14; confirm identity with antibody controls |
| Band near 186.5 kDa in a nuclear fraction | Consistent with KIF14's reported nuclear localization |
| Band near 186.5 kDa in a cytoplasmic fraction | Consistent with KIF14's reported cytoplasmic localization |
| Close doublet near 186.5 kDa | Could reflect phosphorylation, but distinct migration is unverified |
💡Expected KIF14 appearanceUniProt predicts 186.5 kDa for full-length KIF14; no empirical band size is supplied, so confirm any band near that position with ordinary identity controls.
| UniProt predicted molecular weight | Places full-length KIF14 near 186.5 kDa |
| Full-length sequence mass | Provides a 186,492 Da reference for band position |
| 1648-residue protein mass | Supports the predicted full-length band position near 186.5 kDa |
| Predicted full-length reference size | Does not establish an experimentally observed migration position |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KIF14 signal may be below detection in the sampled lysate | Check loading and antibody performance with a positive-control lysate |
| Band higher than expected | The cause is not established by the supplied features | Compare with a positive control and check antibody specificity |
| Band lower than expected | The cause is not established by the supplied features | Check sample integrity and confirm the band with a second antibody |
| Multiple bands | Phosphorylation is documented, but distinct migrating forms are unverified | Compare phosphatase-treated and untreated samples and confirm antibody specificity |
| Weak or no signal | KIF14 may be scarce in the sampled fraction | Check loading and compare cytoplasmic and nuclear fractions with appropriate controls |
Experiment evidence
Western blot images showing KIF14 band
A04137 · KIF14
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KIF14 Western blot
🧪For positive controls for KIF14 in Western blot, you can use a validated KIF14-positive sample; the supplied HPA data identify no specific 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 the samples.
⚠️Feasibility: With no HPA expression data, tissue controls cannot be selected; a KIF14 knockdown or KO line can provide a negative control.
HPA tissue expression evidence for KIF14
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 KIF14 Western Blot Tips
Deeper troubleshooting and optimisation questions for KIF14, answered from its protein features.
Where should the main KIF14 band appear?
Band shift · KIF14 has a predicted mass of 186.5 kDa. Use that as a reference when identifying the band; no observed band position is supplied. Its annotated phosphorylation sites do not establish a visible shift or explain any difference from the calculated mass.
Could KIF14 isoforms explain multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. Do not assign additional bands to KIF14 isoforms from these features alone.
Which KIF14 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at 12, 272, 346, 937 and 1292, and phosphothreonine at 277 and 915. These are UniProt sequence coordinates; paper or antibody numbering may differ. Their presence alone does not establish a visible band shift.
Does this guide establish induction of KIF14?
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 KIF14 Western blot?
Transfer · For a 186.5 kDa protein, use transfer conditions suitable for large proteins, such as a sufficiently long wet transfer. Check transfer with a total-protein stain, especially near the expected KIF14 position.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04137 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 KIF14 localization affect quantitation?
Quantitation · KIF14 is annotated in the nucleus, cytoplasm, spindle and midbody. Compare like samples and fractions, and normalize within the material loaded. A change in one fraction may reflect redistribution rather than a change in total KIF14.
How should unexpected KIF14 bands be interpreted?
Interpretation · First compare them with the 186.5 kDa predicted mass and verify antibody specificity. The supplied features list phosphorylation and one isoform, but neither establishes the identity or cause of an unexpected band. No empirical KIF14 band position is available here.
Boster reagents
KIF14 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-Kinesin-like protein KIF14 KIF14 Antibody
Cat # A04137
The catalog reports A04137 for KIF14 Western blotting, with reported human, mouse, and rat reactivity. Its WB image uses extracts from various cell lines at 1:1000 dilution; the supplied caption does not identify the cell lines or establish validation across all three species.
Which to pick: A04137 is the only listed option and has a WB image. Check that its reported reactivity covers your species, and treat the cell-line image as evidence for the tested conditions rather than every sample type.
Source: BosterBio KIF14 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15058.
- Human Protein Atlas. KIF14 tissue expression.
- PMC3953446 — target-verified WB comparison
- PMC4044675 — target-verified WB comparison
- PMC7078655 — target-verified WB comparison