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KIF15 / Kinesin-like protein KIF15 · IHC design guide
Design Immunohistochemistry for KIF15
This guide pairs KIF15 tissue staining patterns with a paraffin-section IHC workflow (HPA tissue IHC; datasheet A05983-1). Use colon glandular or testis Leydig cells as positive references (HPA tissue IHC), and start the catalog antibody at 0.5–1 µg/mL (datasheet A05983-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KIF15 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 and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining across several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05983-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding was observed and disregarded (HPA tissue IHC) | |
| Regulation | RNA group-enriched in lymphoid tissue (HPA tissue IHC: RNA) | |
| Isoform / epitope | 4 isoforms; map the epitope; no extracellular domain (UniProt) |
Section 1
Recommended KIF15 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is supplemented by published KIF15 IHC conditions for human hepatocellular carcinoma biopsies (PMC7157793) and tissue sections (PMC11909966).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A05983-1) |
| Fixation | Image fixative and duration unreported (datasheet A05983-1); 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 A05983-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05983-1) |
| Primary antibody | Rabbit anti-KIF15, 0.5-1μg/ml (datasheet A05983-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05983-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05983-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KIF15-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A05983-1), then optimize the primary dilution using the published 1:50 and 1:200 conditions (PMC7157793; PMC11909966).
Section 2
What Is the Expected KIF15 Staining Pattern?
KIF15 is a cytoplasmic motor that appears as fine puncta in interphase and on the mitotic spindle during division (UniProt Q9NS87). In tissue IHC, expect staining in selected glandular, germinal center, Leydig, cortical thymic, and hematopoietic cells (HPA tissue IHC). HPA also reports nuclear staining in several tissues; its tissue IHC assessment is Approved, with presumed off-target binding disregarded and external verification pending (HPA tissue IHC). KIF15 has no transmembrane segment (UniProt Q9NS87 topology).
| Cytoplasmic staining in colon glandular cells or lymph-node germinal center cells, with some nuclear staining. | This fits two HPA high-staining cell populations and its broader cytoplasmic and nuclear tissue profile (HPA tissue IHC). Cytoplasmic signal is consistent with interphase KIF15 (UniProt Q9NS87). Score the named cell population and compartment separately; nuclear staining alone cannot establish specificity because HPA reports possible off-target binding (HPA tissue IHC). |
| A spindle-shaped signal in an identifiable mitotic cell. | Spindle localization agrees with KIF15's role in mitotic spindle assembly (UniProt Q9NS87). Interpret the signal alongside cell morphology and surrounding tissue staining; a chromogenic section may not resolve the fine spindle pattern clearly (general IHC practice). A single spindle-like profile is insufficient to validate the whole stain (general IHC practice). |
| Predominantly continuous cell-surface or extracellular staining, without the expected cellular pattern. | This is atypical for KIF15's reported cytoplasmic and spindle localization and lack of a transmembrane segment (UniProt Q9NS87). Review tissue morphology, detection controls, and antibody specificity before calling it KIF15; division-site membrane association is reported only by similarity and does not establish a general surface pattern (UniProt Q9NS87). |
| Strong signal in adipocytes or esophageal squamous epithelial cells. | Both are listed as not detected in HPA tissue IHC, so widespread staining there warrants scrutiny (HPA tissue IHC). Compare a no-primary control for endogenous detection activity and examine whether signal follows cell boundaries or deposits nonspecifically (general IHC practice). HPA's Approved assessment also notes presumed off-target binding (HPA tissue IHC). |
| No staining in colon glandular cells or testis Leydig cells. | Both are high-staining reference populations in HPA tissue IHC, making an entirely blank positive-control section a reason to check the run (HPA tissue IHC). Verify tissue identity, antibody and detection steps, and the control slide before interpreting a study specimen as negative (general IHC practice). HPA levels describe observed patterns, not guaranteed results under every assay condition (HPA tissue IHC). |
💡Expected KIF15 appearanceCall a result convincing when identifiable high-staining populations, such as colon glandular or lymph-node germinal center cells, show cellular cytoplasmic staining with possible nuclear staining (HPA tissue IHC), while uniform extracellular or widespread staining in HPA not-detected populations raises concern (UniProt Q9NS87; HPA tissue IHC).
| Tissue and cell selection | Use the named cell population, not the whole organ, to judge a control: colon glandular, lymph-node germinal center, and testis Leydig cells are High; bone-marrow hematopoietic cells are Medium (HPA tissue IHC). Adipocytes and esophageal squamous epithelial cells are listed as not detected (HPA tissue IHC). |
| Cell state and compartment | Interphase KIF15 is reported as fine cytoplasmic puncta and irregular dots; mitotic KIF15 is reported on the spindle (UniProt Q9NS87). HPA's tissue IHC profile also includes nuclear staining, so nuclear signal requires context rather than automatic rejection (HPA tissue IHC). |
| Antibody evidence | Three listed HPA rabbit polyclonal antibodies have Approved IHC status, with no Enhanced designation supplied (HPA antibodies). The HPA tissue assessment notes presumed off-target binding, supporting data, and pending external verification; retain appropriate tissue and detection controls when judging an unfamiliar pattern (HPA tissue IHC; general IHC practice). |
| Isoforms and epitope scope | UniProt lists four KIF15 isoforms and a kinesin motor domain at residues 26–363 (UniProt Q9NS87). No antibody epitope is supplied, so the record cannot establish which isoforms a given IHC stain detects or whether a particular retrieval condition exposes its epitope (UniProt Q9NS87; HPA antibodies). |
| IF/ICC Q: What pattern is supported here? | A: UniProt reports interphase cytoplasmic puncta and mitotic spindle localization (UniProt Q9NS87). HPA supplies no main ICC-IF location or cell-line images, and the listed antibodies have no ICC status supplied; use those statements as a localization expectation, not as ICC validation (HPA subcellular; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive control is blank. | A failed antibody or detection step is possible; colon glandular and testis Leydig cells are reported High (HPA tissue IHC; general IHC practice). | Check that the control contains the specified cells, then review antibody application, chromogen, and counterstain against the run record (general IHC practice). Avoid calling study sections negative until the positive control works (general IHC practice). |
| The slide shows diffuse haze across cells and extracellular space. | Background from detection chemistry or inadequate blocking can obscure cellular localization (general IHC practice). Diffuse extracellular staining does not match the reported cytoplasmic and spindle pattern (UniProt Q9NS87). | Inspect a no-primary control, review blocking and wash steps, and compare cell-resolved staining with the HPA tissue pattern (general IHC practice; HPA tissue IHC). |
| Nuclear staining dominates without clear cytoplasmic signal. | HPA reports nuclear as well as cytoplasmic tissue staining, while UniProt emphasizes cytoplasmic and spindle localization; nuclear signal alone is therefore inconclusive (HPA tissue IHC; UniProt Q9NS87). | Check whether the named high-staining cells also show a coherent cellular pattern, then compare controls and another listed IHC-Approved antibody if available (HPA tissue IHC; HPA antibodies; general IHC practice). |
| A not-detected reference cell population stains strongly. | HPA lists adipocytes and esophageal squamous epithelial cells as not detected and acknowledges presumed off-target binding in its tissue assessment (HPA tissue IHC). Endogenous detection activity is another general IHC possibility (general IHC practice). | Confirm the cell identity and run a no-primary detection control; interpret persistent signal cautiously alongside an HPA high-staining population (general IHC practice; HPA tissue IHC). |
| Only a few mitotic cells show spindle-associated staining. | Spindle localization is tied to mitosis, whereas interphase localization is cytoplasmic and punctate (UniProt Q9NS87). The number of mitotic cells visible depends on the section sampled (general IHC practice). | Examine identifiable interphase cells in an HPA high-staining population and score their cellular staining separately from spindle profiles (HPA tissue IHC; UniProt Q9NS87; general IHC practice). |
| Results differ between two tissue sections. | HPA reports High, Medium, Low, and not-detected staining across specific cell populations, so different tissue composition can change the apparent result (HPA tissue IHC). | Compare the same named cell type and compartment across sections, record the control outcome, and avoid treating whole-section intensity as a cell-specific score (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing KIF15 staining
A05983-1 · IHCIHC analysis of KIF15 using anti-KIF15 antibody (A05983-1). KIF15 was detected in paraffin-embedded section of human colon cancer tissue. He
A05983-1 · IHCIHC analysis of KIF15 using anti-KIF15 antibody (A05983-1). KIF15 was detected in paraffin-embedded section of rat testis tissue. Heat media
A05983-1 · IFIF analysis of KIF15 using anti-KIF15 antibody (A05983-1). KIF15 was detected in immunocytochemical section of A431 cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163808-KIF15/tissue/cerebral+cortex#imid_20852911
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163808-KIF15/tissue/colon#imid_20852845
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163808-KIF15/tissue/liver#imid_20852854
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163808-KIF15/tissue/kidney#imid_20852855
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KIF15 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the colon slide, assess non-glandular cells for background staining rather than assuming they lack KIF15.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for KIF15; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and matched rabbit IgG isotype controls (caption: rabbit anti-KIF15 primary), plus KIF15 knockout material or a validated peptide-block control. For colon sections developed with biotin-based DAB detection (caption: biotinylated secondary and SABC/DAB), check endogenous peroxidase and biotin background.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A05983-1 tissue-IHC caption does not state the fixative. That paraffin-section example used heat retrieval in EDTA at pH 8.0 (caption: EDTA retrieval); optimize retrieval for the available specimen without assuming a paraffin-section result. IF may help assess cytoplasmic puncta and mitotic spindle localisation (UniProt Q9NS87: subcellular location), but the supplied HPA record has no ICC-IF images (HPA: subcellular record), so relative ease of frozen-section or IF staining is unreported. The selected A05983-1 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A05983-1).
HPA tissue IHC evidence for KIF15
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Presumed off target binding observed and disregarded. External characterization data supports antibody staining. Pending external verification.). 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KIF15 IHC Tips
Troubleshoot KIF15 staining in paraffin sections by checking retrieval, cellular pattern, controls and scoring before interpreting chromogenic signal.
How should I retrieve KIF15 in paraffin sections when staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A05983-1). The selected paraffin-section example used this retrieval before 1 μg/ml primary antibody overnight at 4°C, so reproduce those conditions when comparing runs (datasheet A05983-1). If staining remains weak, check that sections were fully deparaffinised and that heating, cooling and wash steps were consistent across slides; change one variable at a time and retain a positive control (standard IHC practice). Record retrieval duration and temperature locally, because the selected caption specifies the buffer and pH but gives no heating time or temperature (datasheet A05983-1).
Could fixation explain variable KIF15 staining between paraffin blocks?
It could affect antigen access, but KIF15-specific fixation sensitivity is unknown from the supplied evidence (datasheet A05983-1: fixative unreported). The selected image documents a paraffin-embedded colon cancer section and pH 8.0 EDTA retrieval; it does not establish formalin fixation or a preferred fixation duration (datasheet A05983-1). Compare blocks with documented processing histories using the same retrieval, 1 μg/ml primary concentration and detection workflow, then inspect morphology alongside signal (datasheet A05983-1; standard IHC practice). If one block fails, include a positive tissue section in the same run before attributing the difference to biology; do not infer fixation sensitivity from expression or protein structure (standard IHC practice).
What KIF15 pattern should I expect in chromogenic tissue sections?
Assess staining by compartment and cell type rather than treating every brown deposit as KIF15 (standard IHC practice). KIF15 is described as cytoplasmic, finely punctate during interphase and associated with the mitotic spindle during mitosis (UniProt Q9NS87: subcellular location). Its spindle association may be hard to resolve in routine chromogenic sections, while the tissue atlas reports cytoplasmic and nuclear staining across several tissues (UniProt Q9NS87: subcellular location; HPA: tissue profile). Document nuclear and cytoplasmic signal separately, compare it with morphology and controls, and avoid calling nuclear staining specific solely because it appears in the atlas (HPA: presumed off-target binding; standard IHC practice).
How do KIF15 isoforms and epitope location affect a negative stain?
KIF15 has 4 annotated isoforms and a kinesin motor domain at residues 26–363 (UniProt Q9NS87: isoforms and domains). The supplied antibody caption does not identify its immunogen or mapped epitope, so a negative section cannot establish which isoforms are absent (datasheet A05983-1: caption). Check the antibody's epitope documentation before making isoform-specific claims, and compare staining with an independently characterised antibody if isoform coverage matters (standard IHC practice). Report the antibody identity and retrieval conditions with the result; phosphorylation and acetylation sites are annotated, but their effects on this antibody's staining are unestablished (UniProt Q9NS87: modified residues; datasheet A05983-1: caption).
How should I use IF to investigate an ambiguous KIF15 IHC pattern?
Use IF as an orthogonal localisation check while keeping the paraffin-section IHC result as the primary readout (standard IHC/IF practice). Multiplex KIF15 with a marker for the cell population being evaluated, and use separate channels plus single-stain controls to distinguish overlap from spectral bleed-through (standard IF practice). Select a fluorophore channel with low tissue autofluorescence, verified on an unstained section, before comparing its pattern with chromogenic signal (standard IF practice). KIF15 lacks a transmembrane segment and is reported in the cytoplasm and spindle, so permeabilise for access to intracellular epitopes; optimise that step for the IF preparation and antibody (UniProt Q9NS87: topology and localisation; standard IF practice).
How can I reduce diffuse brown background without losing KIF15 signal?
First compare the stained section with a no-primary control to separate detection-system background from primary-antibody-associated signal (standard IHC practice). The selected example used 10% goat serum, 1 μg/ml primary overnight at 4°C, a biotinylated secondary and DAB development (datasheet A05983-1). Keep those conditions documented while checking washing, serum blocking and DAB development, and include a peroxidase block as a general chromogenic IHC step (standard IHC practice). Because the selected detection uses a biotin-based complex, investigate endogenous biotin if background follows tissue structures in the no-primary control; do not classify uniform staining as KIF15 from colour alone (datasheet A05983-1; standard IHC practice).
How should I quantify KIF15 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then apply the same rule to every section (standard IHC practice). For chromogenic signal, record percentage of positive cells and intensity grades, or calculate an H-score from 0–300; keep nuclear and cytoplasmic scores separate when both appear (standard IHC practice; HPA: tissue profile). Normalise positive-cell counts to the number of eligible cells, or report density per mm² of viable tissue when cell counting is impractical (standard IHC practice). Exclude necrosis, folds and tissue edges by a prespecified rule, and run comparable sections together with control tissue to limit processing and staining variation (standard IHC practice).
When is a positive KIF15 stain convincing rather than artefactual?
Look for reproducible staining in intact cells, with a plausible cytoplasmic distribution and appropriate controls, before calling a section positive (UniProt Q9NS87: localisation; standard IHC practice). The atlas reports high glandular-cell staining in colon and high germinal-center-cell staining in lymph node, but also flags presumed off-target binding, so atlas agreement alone does not prove specificity (HPA: tissue IHC). Question signal confined to an implausible compartment or cell population, section edges or necrotic areas, and compare it with no-primary and positive controls (standard IHC practice). Brown precipitate attributable to endogenous peroxidase or the detection system should be resolved before interpreting differences in KIF15 expression (standard IHC practice).
Boster reagents
Best KIF15 / Kinesin-like protein KIF15 IHC Antibodies
A05983-1 has IHC images from human colon cancer and rat testis paraffin sections, plus an IF image from A431 cells (catalog image captions).
Real IHC data
Anti-KIF15 Antibody ®
Cat # A05983-1
A05983-1 was used for IHC on human colon cancer and rat testis paraffin sections (catalog IHC image captions). The same SKU was used for IF in A431 cells (catalog IF image caption).
Which to pick: For tissue IHC, choose A05983-1: its images show paraffin sections with EDTA retrieval at pH 8.0, and the fixative is unreported (catalog IHC image captions). For IF/ICC, choose A05983-1; IF and ICC are listed applications, and its A431 IF image used 5 μg/mL (catalog applications; catalog IF image caption). For human and rat work, A05983-1 lists both species as reactive, with IHC images for each; it is rabbit hosted, while clonality is unreported (catalog reactivity and host; catalog IHC image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NS87 (KIF15_HUMAN, Kinesin-like protein KIF15).
- Human Protein Atlas. KIF15 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KIF15 subcellular location (ICC-IF): Highest expression in AF22: 16.5 nTPM.
- Human Protein Atlas. KIF15 antibody validation summary (3 antibodies).
- KIF15 Promotes Proliferation and Growth of Hepatocellular Carcinoma. Analytical cellular pathology (Amsterdam) 2020 — PMC7157793.
- Atractylenolide I ameliorated the growth and enzalutamide resistance of castration-resistant prostate cancer by targeting KIF15. Chinese medicine 2025 — PMC11909966.
- Dynamic regulation of KIF15 phosphorylation and acetylation promotes focal adhesions disassembly in pancreatic cancer. Cell death & disease 2022 — PMC9592618.
- PSMD12 promotes the activation of the MEK-ERK pathway by upregulating KIF15 to promote the malignant progression of liver cancer. Cancer biology & therapy 2022 — PMC9519003.
- PubMed PMID:10878014 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KIF15 (Kinesin-like protein KIF15). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05983-1). Validated IHC antibody: A05983-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A05983-1); 2. Fixation: Image fixative and duration unreported (datasheet A05983-1); 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 A05983-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05983-1); 8. Primary antibody: Rabbit anti-KIF15, 0.5-1μg/ml (datasheet A05983-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05983-1). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Presumed off-target binding was observed and disregarded (HPA tissue IHC); Isoform / epitope: 4 isoforms; map the epitope; no extracellular domain (UniProt); Regulation: RNA group-enriched in lymphoid tissue (HPA tissue IHC: RNA). Evidence: UniProt Q9NS87, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in AF22: 16.5 nTPM, Datasheet A05983-1, PMC7157793, PMC11909966. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KIF15/immunohistochemistry-ihc-guide.