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KIF11 / Kinesin-like protein KIF11 · IHC design guide
Design Immunohistochemistry for KIF11
Plan KIF11 chromogenic IHC in paraffin sections using the catalog antibody at 2–5 μg/mL (datasheet A01754-2). Compare cytoplasmic staining across the cell subsets highlighted by tissue IHC evidence (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KIF11 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in tissue (HPA tissue IHC); spindle-pole localisation is a molecular annotation (UniProt) | |
| Staining pattern | Cytoplasmic in seminiferous, gut and lymphoid cell subsets (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01754-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Caudate |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01754-2) | |
| Caveat | Staining varies between cell subsets (HPA tissue IHC) | |
| Regulation | Expression regulation unannotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–1056 chain (UniProt) |
Section 1
Recommended KIF11 IHC & IF Protocols
The catalog antibody protocol is followed by three published KIF11 IHC protocols (PMC11898751; PMC5696167; PMC5696720).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01754-2) |
| Fixation | Image fixative and duration unreported (datasheet A01754-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A01754-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01754-2) |
| Primary antibody | Rabbit anti-KIF11, 2-5 μg/ml (datasheet A01754-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01754-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01754-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KIF11-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. Highest levels in subsets of cells in seminiferous duct, the gastrointestinal tract, in lymphoid tissue and bone marrow. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01754-2); optimize staining on the study tissue.
Section 2
What Is the Expected KIF11 Staining Pattern?
KIF11 is a cytoplasmic motor associated with the spindle pole and has no transmembrane segment (UniProt P52732). In paraffin section IHC, expect cytoplasmic staining in subsets of cells, including glandular cells of the colon and duodenum, lymph node germinal center cells, and bone marrow hematopoietic cells (HPA: High in each). HPA rates its tissue IHC profile Enhanced, while describing medium consistency between staining and RNA expression (HPA: tissue IHC).
| Cytoplasmic staining is strongest in a subset of glandular, lymphoid, or hematopoietic cells. | This fits the reported compartment and uneven tissue distribution (UniProt P52732: cytoplasm; HPA: cytoplasmic expression in several tissues). Compare cells within the same section: HPA reports High staining in colon glandular cells, lymph node germinal center cells, and bone marrow hematopoietic cells, without implying that every cell must stain (HPA: tissue IHC). |
| Strong staining is predominantly nuclear or confined to a cell membrane, with little cytoplasmic signal. | Treat the compartment mismatch as an unconfirmed result and check controls, morphology, and detection background (UniProt P52732: cytoplasm and spindle pole; no transmembrane segment). A mitotic spindle related pattern may occur in dividing cells, but a nuclear or membrane dominant pattern alone does not match the stated localization (UniProt P52732: function and subcellular location). |
| Cells expected to be weak or negative stain as strongly as the expected positive population. | Check for cross reactivity or endogenous chromogenic detection activity before interpreting this as KIF11 (general IHC practice). For a concrete comparison, HPA reports KIF11 as Not detected in caudate glial cells, while reporting High staining in lymph node germinal center cells (HPA: tissue IHC); these observations are tissue and cell specific. |
| Diffuse color covers extracellular space, several compartments, or most cells without a clear cellular pattern. | This is less convincing than cell associated cytoplasmic staining (UniProt P52732: cytoplasm; HPA: tissue IHC profile). Review the no primary control, blocking, washes, and chromogen development to assess background (general chromogenic IHC practice). Background alone cannot establish KIF11 expression or a target specific fixation effect. |
| A section chosen for an HPA High population has no visible signal in that population. | First confirm that the relevant cells are present and identifiable; HPA levels refer to specified cell populations, not an entire organ (HPA: tissue IHC). If morphology is adequate, review primary antibody application, retrieval conditions, detection reagents, and the positive control (general IHC practice). The supplied sources do not identify a KIF11 specific cause of signal loss. |
💡Expected KIF11 appearanceCall a positive result when identifiable glandular, germinal center, or hematopoietic cell subsets show clear cytoplasmic chromogen above background (HPA: High in colon, lymph node, and bone marrow; UniProt P52732: cytoplasm); diffuse color or dominant nuclear or membrane staining is an unconfirmed pattern requiring control review (general IHC practice; UniProt P52732: localization and topology).
| Compartment and cell cycle | KIF11 is annotated in the cytoplasm and at the spindle pole, and its motor function supports bipolar spindle formation during mitosis (UniProt P52732). Assess staining against cellular morphology and avoid requiring a visible spindle in every positive cell; KIF11 also has a reported non mitotic transport function (UniProt P52732). |
| Cell population within the section | HPA describes cytoplasmic expression in several tissues and its highest levels in subsets of cells (HPA: tissue IHC profile). Score the named cell population rather than averaging across the section: colon glandular cells, lymph node germinal center cells, and bone marrow hematopoietic cells are each High, whereas caudate glial cells are Not detected (HPA: tissue IHC). |
| Strength of IHC evidence | HPA calls the tissue profile Enhanced but describes medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). Its antibody records differ: HPA010568 is IHC Enhanced, while HPA006916 and CAB017617 are IHC Supported (HPA: antibody validation). Interpret an unexpected compartment or cell population with the validation status of the antibody actually used. |
| Antigen retrieval and detection workflow | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in identifiable colon or duodenal glandular cells, or lymph node germinal center cells. | A failed assay step or unsuitable control section is possible; these populations are reported High, but staining is not specified for every cell (HPA: tissue IHC; general IHC practice). | Verify morphology and run a known positive section alongside a no primary control; then check primary antibody application, retrieval, and detection steps (general IHC practice). Do not assign the failure to KIF11 fixation sensitivity: it is unreported in the supplied sources. |
| Signal appears mainly in nuclei or along plasma membranes. | The dominant compartment conflicts with the annotated cytoplasm and spindle pole and with the absence of a transmembrane segment (UniProt P52732). | Recheck cell boundaries and the no primary control, then compare with a documented cytoplasmic positive population (general IHC practice; HPA: High in colon glandular cells). Treat the unusual localization as unresolved unless independently supported. |
| Caudate glial cells stain strongly. | HPA reports KIF11 as Not detected in caudate glial cells; cross reactivity or background is possible, though a single discrepant section cannot identify the cause (HPA: tissue IHC; general IHC practice). | Confirm glial cell identity and compare the same run with an HPA High cell population and a no primary control (HPA: tissue IHC; general IHC practice). Reassess antibody specificity if strong cell associated staining persists. |
| Chromogen is widespread and obscures cell boundaries. | Excess detection background or endogenous activity can create color unrelated to the expected cytoplasmic distribution (general chromogenic IHC practice; UniProt P52732: cytoplasm). | Inspect no primary and detection controls, then review blocking, washes, and chromogen development (general IHC practice). Score KIF11 only where signal remains clearly cell associated and exceeds background. |
| Only scattered cells are positive in a tissue expected to stain. | Scattered staining may reflect the reported subset pattern rather than assay failure; HPA describes its highest levels in subsets of cells (HPA: tissue IHC profile). | Identify and score the relevant population, then compare its cytoplasmic signal with nearby cells and controls (HPA: tissue IHC; general IHC practice). Do not convert a population specific HPA level into a requirement for uniform staining. |
| What pattern should an IF/ICC image show? | HPA reports mainly cytosol and mid piece localization; it also lists mitotic spindle localization as uncertain (HPA: subcellular ICC-IF). These image annotations are distinct from its tissue IHC observations. | For IF/ICC interpretation, look first for the supported cytosol or approved mid piece pattern and treat spindle staining cautiously (HPA: subcellular ICC-IF). Use the separate IF/ICC guide for experimental conditions. |
Experiment evidence
IHC & IF images showing KIF11 staining
A01754-2 · IHCIHC analysis of Eg5/KIF11 using anti-Eg5/KIF11 antibody (A01754-2). Eg5/KIF11 was detected in a paraffin-embedded section of human breast ca
A01754-2 · IHCIHC analysis of Eg5/KIF11 using anti-Eg5/KIF11 antibody (A01754-2). Eg5/KIF11 was detected in a paraffin-embedded section of human endometri
A01754-2 · IFIF analysis of Eg5/KIF11 using anti-Eg5/KIF11 antibody (A01754-2). Eg5/KIF11 was detected in an immunocytochemical section of A431 cells. En
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138160-KIF11/tissue/cerebral+cortex#imid_3232760
HPA · Oral mucosaHuman Protein Atlas IHC, Oral mucosa. Source: https://www.proteinatlas.org/ENSG00000138160-KIF11/tissue/oral+mucosa#imid_3232676
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138160-KIF11/tissue/colon#imid_3232686
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138160-KIF11/tissue/liver#imid_3232687
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA010093; green = KIF11, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA010093; green = KIF11, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KIF11 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use caudate glial cells as the negative tissue comparator (HPA: Not detected in caudate glial cells); within the colon section, cells verified as unstained should retain counterstain without specific brown DAB signal.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KIF11 in A-431, Rh30, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Cytosol (supported), Mid piece (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and species- and clonality-matched isotype controls, using rabbit IgG to match the primary in the selected caption (caption: rabbit anti-KIF11); use KIF11 knockout material as a biological negative. Quench endogenous peroxidase and check for intrinsic pigment before interpreting DAB in colon (standard chromogenic IHC practice).
⚠️Feasibility: The selected paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but reports no fixative or target-specific fixation window; the effect of fixation on KIF11 staining is unreported (caption: EDTA retrieval; fixative not stated). Start with that retrieval condition for the catalog antibody and assess the controls alongside it (caption: paraffin-section IHC). Frozen-section performance and whether IF is easier are unreported in the supplied application evidence; the supplied HPA IF images show subcellular localization, not a matched comparison with paraffin IHC (HPA: subcellular ICC-IF images).
HPA tissue IHC evidence for KIF11
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| 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 → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KIF11 IHC Tips
Troubleshoot KIF11 staining in paraffin sections by checking retrieval, cellular localisation, controls, and scoring before interpreting chromogenic signal.
What retrieval should I use if KIF11 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A01754-2). The selected KIF11 tissue image used that retrieval before overnight incubation with 2 μg/ml primary antibody at 4°C (datasheet A01754-2). Run a known positive section alongside the test section, and compare staining in the expected cell population before changing retrieval conditions (HPA: high in bone marrow hematopoietic cells; standard IHC practice). If staining remains weak, test a different retrieval buffer as a fallback on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Avoid judging retrieval from diffuse edge staining or damaged morphology (standard IHC practice).
How can I assess whether fixation is limiting KIF11 detection?
Target-specific fixation sensitivity is unknown: the selected tissue caption identifies a paraffin section but does not state its fixative (datasheet A01754-2). Record the fixative, fixation duration, and processing history for each specimen before comparing staining intensity (standard IHC practice). Compare sections processed together, using the same EDTA pH 8.0 retrieval and antibody conditions, to separate specimen variation from staining variation (datasheet A01754-2; standard IHC practice). If a specimen stains weakly, inspect morphology and include a known positive section in the same run (standard IHC practice). Do not infer a KIF11-specific fixation effect from its tissue distribution or intracellular location (HPA: tissue IHC profile; UniProt P52732 localisation).
Which staining pattern supports KIF11 localisation in tissue?
Expect mainly cytoplasmic signal in tissue sections, with cell-to-cell variation across the section (HPA: cytoplasmic expression in several tissues). KIF11 is also associated with the cytoskeleton and spindle pole, consistent with its role in establishing a bipolar mitotic spindle (UniProt P52732 localisation and function). Examine high-power fields for staining in intact cells, and use morphology to distinguish cytoplasmic signal from pigment or extracellular deposit (standard IHC practice). HPA reports high staining in lymph-node germinal-center cells and bone-marrow hematopoietic cells, which can guide positive-control selection (HPA: lymph node and bone marrow). Treat isolated strong nuclear staining without convincing cytoplasm as a localisation discrepancy requiring controls (UniProt P52732 localisation; standard IHC practice).
Could an isoform or modified epitope explain discordant KIF11 staining?
The supplied record lists 0 annotated isoforms and a single 1–1056 chain, so an isoform-specific explanation is unsupported here (UniProt P52732 isoforms and processing). KIF11 contains a kinesin motor domain at residues 18–359 and reported modified residues including phosphorylated threonines 925–926 (UniProt P52732 domains and modified residues). The antibody epitope is not specified in the supplied caption, so do not assign weak tissue staining to any particular modification (datasheet A01754-2 caption). Compare staining under the documented EDTA pH 8.0 retrieval with a positive section in the same run (datasheet A01754-2; standard IHC practice). If a second antibody is available, require independent epitope information before treating discordance as biological (standard IHC practice).
How should I follow up an ambiguous chromogenic pattern with IF?
Use IF as a localisation check for ambiguous IHC signal, pairing KIF11 with a validated marker for the expected cell population, such as a germinal-center cell marker in lymph node (HPA: high in lymph-node germinal-center cells; standard IF practice). Choose a fluorophore in a channel with low tissue autofluorescence and inspect single-channel controls before interpreting overlap (standard IF practice). KIF11 has no transmembrane segment and is reported in the cytoplasm, so permeabilise sufficiently to expose an intracellular epitope while preserving cellular structure (UniProt P52732 topology and localisation; standard IF practice). HPA reports cytosol as a supported IF location and mitotic spindle as uncertain, so grade those observations accordingly (HPA: subcellular localisation). Keep the tissue IHC result and IF observation traceable to their respective preparations (standard practice).
What should I check when KIF11 DAB staining looks diffuse?
First inspect a no-primary control for secondary-reagent binding and chromogen deposition, then check the peroxidase block used for the chromogenic run (standard IHC practice). The selected image used a peroxidase-conjugated anti-rabbit secondary and DAB, but its caption does not establish a KIF11-specific background mechanism (datasheet A01754-2). Review blocking and washes before increasing primary antibody: the documented section received 10% goat-serum block and 2 μg/ml primary overnight at 4°C (datasheet A01754-2). Compare diffuse staining with cellular morphology and an expected positive population, rather than scoring stained debris (HPA: tissue IHC profile; standard IHC practice). If background persists, vary one staining step at a time on adjacent sections (standard IHC practice).
How should I quantify KIF11 across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring compartment before reviewing outcomes, since HPA describes cytoplasmic expression that varies among tissues and cell subsets (HPA: tissue IHC profile). For a defined population, report the percentage of positive cells and an H-score using intensity categories 0–3; for focal populations, report positive-cell density per mm² of viable tissue (standard IHC practice). Normalise counts to the number of evaluable cells or viable tissue area, and record excluded necrotic or damaged regions (standard IHC practice). Apply the same threshold, counterstain conditions, and field-selection rule across specimens (standard IHC practice). Report control-section performance with the scores because HPA assigns the tissue profile Enhanced reliability with medium staining–RNA consistency (HPA: tissue IHC reliability).
How do I distinguish a true KIF11 positive from artefact?
A credible positive is cell-associated and predominantly cytoplasmic in an intact, plausible population, with appropriate control staining (HPA: tissue IHC profile; standard IHC practice). KIF11 also associates with spindle poles during mitosis, so a convincing mitotic pattern should be judged in the context of cell morphology rather than an isolated dark dot (UniProt P52732 localisation and function; standard IHC practice). Question staining confined to section edges, necrotic areas, or extracellular deposits, and use a no-primary control to assess endogenous enzyme or detection background (standard IHC practice). Compare the suspected population with documented high-staining populations, such as lymph-node germinal-center cells, without assuming every cell must stain (HPA: lymph node; HPA: tissue IHC profile). Resolve a compartment mismatch before assigning biological significance (UniProt P52732 localisation; standard IHC practice).
Boster reagents
Best KIF11 / Kinesin-like protein KIF11 IHC Antibodies
A01754-2 has IHC data from human paraffin sections and IF/ICC data from A431 cells (A01754-2 image captions); its listed reactivity is human (catalog: A01754-2 reactivity).
Real IHC data
Anti-Eg5/KIF11 Antibody ®
Cat # A01754-2
A01754-2 will render with its human breast cancer paraffin-section IHC figure (A01754-2 IHC caption). The same SKU has an A431 cell IF image (A01754-2 IF caption).
Which to pick: Choose A01754-2 for human paraffin-section IHC: its own caption documents EDTA pH 8 retrieval and chromogenic detection, but does not report the fixative (A01754-2 IHC caption). For IF/ICC, A01754-2 has A431 image data; M01754-3 is a rabbit monoclonal listed for human ICC/IF without a supplied image (A01754-2 IF caption; catalog: M01754-3 host, clone, applications, reactivity, IF image alts). Neither SKU has stated reactivity beyond human, so the payload supports no cross-species recommendation (catalog: both reactivity fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P52732 (KIF11_HUMAN, Kinesin-like protein KIF11).
- Human Protein Atlas. KIF11 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. KIF11 subcellular location (ICC-IF): Mainly localized to the cytosol and mid piece. In addition localized to vesicles, mitotic spindle, basal body, principal piece and end piece..
- Human Protein Atlas. KIF11 antibody validation summary (4 antibodies).
- KIF11 and KIF14 Are a Novel Potential Prognostic Biomarker in Patients with Endometrioid Carcinoma. Cancers 2025 — PMC11898751.
- KIF11 is required for proliferation and self-renewal of docetaxel resistant triple negative breast cancer cells. Oncotarget 2017 — PMC5696167.
- Kinesin family member 11 contributes to the progression and prognosis of human breast cancer. Oncology letters 2017 — PMC5696720.
- KIF11 promotes vascular smooth muscle cell proliferation by regulating cell cycle progression and accelerates neointimal formation after arterial injury in mice. Frontiers in pharmacology 2024 — PMC11333341.
- PubMed PMID:8548803 — UniProt-cited evidence.
- PubMed PMID:9701554 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KIF11 (Kinesin-like protein KIF11). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC); spindle-pole localisation is a molecular annotation (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01754-2). Validated IHC antibody: A01754-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01754-2); 2. Fixation: Image fixative and duration unreported (datasheet A01754-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A01754-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01754-2); 8. Primary antibody: Rabbit anti-KIF11, 2-5 μg/ml (datasheet A01754-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01754-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01754-2); Caveat: Staining varies between cell subsets (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–1056 chain (UniProt); Regulation: Expression regulation unannotated (UniProt). Evidence: UniProt P52732, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol and mid piece. In addition localized…, Datasheet A01754-2, PMC11898751, PMC5696167. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KIF11/immunohistochemistry-ihc-guide.