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- Table of Contents
KIF11 / Kinesin-like protein KIF11 · Western blot design guide
Design a Western Blot for KIF11
Real validated KIF11 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KIF11 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
KIF11 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 | ~119.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated KIF11 Western Blot Protocols
The M01754-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Raji cell lysate (catalog M01754-3) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M01754-3; 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 KIF11 Western Blot Band Size?
KIF11 is predicted at 119.2 kDa; listed modifications could affect migration, but no empirical band or demonstrated shift is supplied.
| Single band near 119.2 kDa | consistent with predicted full-length KIF11 size; confirm identity with a specificity control |
| Band in a cytoplasmic fraction | consistent with KIF11 cytoplasmic localization |
| Band in a spindle-pole fraction | consistent with KIF11 spindle-pole localization |
| Nearby doublet around 119.2 kDa | could reflect listed phosphorylation states, but distinct migration is unproven |
💡Expected KIF11 appearanceFull-length KIF11 has a predicted mass of 119.2 kDa; no empirical band size or modification-dependent shift is supplied, so confirm any candidate band with a specificity control.
| UniProt predicted mass | places full-length KIF11 near 119.2 kDa |
| Phosphothreonine at T458 | may affect migration, but no size shift is established |
| Phosphothreonine at T925 and T926 | may affect migration, but no size shift is established |
| Phosphoserine at S1033 | may affect migration, but no size shift is established |
| N6-acetyllysine at K146 | has no established visible size effect |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KIF11 may be below detection in the sampled lysate | check loading and use a validated positive lysate and antibody control |
| Band higher than expected | its identity or modification-dependent migration is unconfirmed | compare with a specificity control and a molecular-weight marker |
| Band lower than expected | a smaller species has no documented cleavage explanation here | check antibody specificity and sample handling |
| Multiple bands | listed phosphorylation sites could contribute, but distinct bands are unproven | compare phosphatase-treated and untreated samples with a specificity control |
| Weak or no signal | the sampled fraction may contain little cytoplasmic or spindle-pole KIF11 | check loading and compare appropriate cellular fractions |
Experiment evidence
Western blot images showing KIF11 band
M01754-3 · KIF11
A01754-2 · KIF11
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KIF11 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KIF11 in Western blot, you can use appendix tissue, which HPA rates as high expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Tissue controls are feasible because KIF11 is intracellular, with high expression in appendix and no detected expression in caudate.
HPA tissue expression evidence for KIF11
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced KIF11 Western Blot Tips
Deeper troubleshooting and optimisation questions for KIF11, answered from its protein features.
How should KIF11 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 a second band represent a KIF11 isoform?
Isoforms · The supplied features list one isoform and no alternative sequence. They do not support assigning a second band to a KIF11 isoform.
Which KIF11 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphothreonine at positions 458, 925 and 926, with CDK1 specified for 926, and phosphoserine at 1033 by NEK6. These are UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish a visible band shift.
UniProt lists N6-acetyllysine at position 146. Use UniProt numbering when comparing this site with antibody or paper annotations. The feature alone does not predict a distinct Western blot band.
UniProt lists N6-acetyllysine at position 146. Use UniProt numbering when comparing this site with antibody or paper annotations. The feature alone does not predict a distinct Western blot band.
Does this guide establish induction of KIF11?
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 KIF11?
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 M01754-3 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.
What should be controlled when quantifying KIF11 bands?
Quantitation · Compare similarly prepared samples and account for KIF11's cytoplasmic and spindle pole localization, especially if using fractions. Normalize loading and keep cell cycle conditions comparable when interpreting differences.
Should KIF11 run at exactly 119.2 kDa?
Interpretation · 119.2 kDa is the predicted mass; no observed band position is supplied. The listed modifications do not, by themselves, establish a visible shift or explain any difference between apparent and predicted mass.
KIF11 is annotated for cell cycle, cell division and mitosis, with localization to the cytoplasm and spindle pole. Record cell cycle conditions when comparing samples; the supplied features do not establish that a particular treatment increases KIF11 abundance.
Compare them with the 119.2 kDa predicted mass, but avoid assigning identities from position alone. The supplied features list one isoform, acetyllysine 146 and four phosphorylation sites; none proves that an extra band is a specific KIF11 form. No empirical band position is supplied.
KIF11 is annotated for cell cycle, cell division and mitosis, with localization to the cytoplasm and spindle pole. Record cell cycle conditions when comparing samples; the supplied features do not establish that a particular treatment increases KIF11 abundance.
Compare them with the 119.2 kDa predicted mass, but avoid assigning identities from position alone. The supplied features list one isoform, acetyllysine 146 and four phosphorylation sites; none proves that an extra band is a specific KIF11 form. No empirical band position is supplied.
Boster reagents
KIF11 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-Eg5 Rabbit Monoclonal Antibody
Cat # M01754-3
Real WB data
Anti-Eg5/KIF11 Antibody Picoband®
Cat # A01754-2
Both listed anti-KIF11 antibodies have Western blot images using human cell lysates. M01754-3 is shown with Raji lysate; A01754-2 is shown with five human cell lines and a reported band near 130 kDa, versus 119 kDa expected. These images document specific tested contexts.
Which to pick: Choose A01754-2 if you want an image covering MCF-7, Raji, 293T, Daudi, and MOLT-4 lysates with reported blot conditions. M01754-3 has an image using Raji lysate. Both list human reactivity.
Source: BosterBio KIF11 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P52732.
- Human Protein Atlas. KIF11 tissue expression.
- PMC9468433 — target-verified WB comparison
- PMC12628542 — target-verified WB comparison
- PMC11730827 — target-verified WB comparison