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KIF2C / Kinesin-like protein KIF2C · IHC design guide
Design Immunohistochemistry for KIF2C
Plan KIF2C chromogenic IHC in paraffin sections using the reported nuclear testis and cytoplasmic lymphoid and intestinal patterns (HPA tissue IHC). Compare cell types within each tissue and interpret staining intensity against those reference patterns (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KIF2C 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 | Nuclear in testis; cytoplasmic in lymphoid tissues and intestines (HPA tissue IHC) | |
| Staining pattern | Nuclear testis staining; cytoplasmic lymphoid and intestinal staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9230) | |
| Positive control | Testis+2 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9230) | |
| Caveat | Staining intensity and compartment vary by tissue (HPA tissue IHC) | |
| Regulation | Highest staining in testis; lower in other sampled tissues (HPA tissue IHC) | |
| Isoform / epitope | Two isoforms; isoform 2 is testis-specific. Check epitope coverage (UniProt) |
Section 1
Recommended KIF2C IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet PB9230) is accompanied by published KIF2C staining methods for Wilms tumor, cervical cancer, and liver tissue (PMC10505208; PMC8828917; PMC9171145).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PB9230); 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 PB9230); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9230) |
| Primary antibody | Rabbit anti-KIF2C, 0.5-1μg/ml (datasheet PB9230) |
| Primary incubation | Overnight at 4 °C (datasheet PB9230) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9230) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KIF2C-positive staining in pachytene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in testis and cytoplasmic expression in lymphoid tissues and intestines. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet PB9230); citrate retrieval is documented for Wilms tumor and cervical cancer sections (PMC10505208; PMC8828917).
Section 2
What Is the Expected KIF2C Staining Pattern?
KIF2C is an intracellular protein with no transmembrane segment (UniProt Q99661 topology). In paraffin-section IHC, expect nuclear staining in testis and cytoplasmic staining in lymphoid tissues and intestines (HPA tissue IHC: Enhanced reliability). Pachytene spermatocytes are a strong positive population; appendix endocrine cells and thymus cortical cells show moderate staining (HPA tissue IHC: High in pachytene spermatocytes; Medium in appendix endocrine and thymus cortical cells).
| Strong nuclear staining in pachytene spermatocytes. | This matches the clearest supplied positive reference (HPA tissue IHC: High in pachytene spermatocytes; nuclear expression in testis). Score the named cell population and compartment together: a positive-looking testis section is less informative if the stained cells cannot be identified. |
| A crisp membrane rim dominates the stained cells. | That pattern is difficult to reconcile with the reported intracellular locations (UniProt Q99661 subcellular location; HPA tissue IHC: nuclear in testis, cytoplasmic in lymphoid tissues and intestines). KIF2C has no transmembrane segment (UniProt Q99661 topology); review the signal before calling it specific. |
| Strong staining appears broadly in a cell population reported as negative. | For example, colon endocrine cells are reported as not detected (HPA tissue IHC: colon endocrine cells). Cross-reactivity or endogenous detection activity is possible; neither can be diagnosed from appearance alone. General IHC practice: compare a no-primary control and a validated positive section. |
| Diffuse colour covers many compartments or extends beyond cell boundaries. | A tissue-wide haze is poorly matched to the cell- and compartment-specific HPA profile (HPA tissue IHC: nuclear in testis; cytoplasmic in lymphoid tissues and intestines). General IHC practice: assess blocking, washing and detection background before scoring faint cellular colour. |
| No staining is seen in identifiable pachytene spermatocytes. | Treat this as a failed positive-control result until checked: these cells are reported as High (HPA tissue IHC: testis pachytene spermatocytes). Confirm that the section contains the relevant cells, then review the antibody's IHC conditions and detection controls (general IHC practice). |
💡Expected KIF2C appearanceCall a result positive when identifiable pachytene spermatocytes show High nuclear staining (HPA tissue IHC: testis); a dominant membrane rim or tissue-wide haze is discordant with the reported intracellular, cell-specific pattern (UniProt Q99661 topology; HPA tissue IHC).
| Cell population and tissue | Expected intensity varies by sampled cell: High in testis pachytene spermatocytes, Medium in appendix endocrine and thymus cortical cells, and Low in several intestinal and lymphoid populations (HPA tissue IHC). Identify cells before comparing sections. |
| Compartment | HPA reports nuclear expression in testis and cytoplasmic expression in lymphoid tissues and intestines (HPA tissue IHC). UniProt also lists cytoskeleton, nucleus, centromere and kinetochore locations (UniProt Q99661 subcellular location); these annotations do not require every feature to resolve in chromogenic IHC. |
| Antibody evidence | The tissue profile has Enhanced reliability (HPA tissue IHC). Two listed antibodies, HPA079172 and CAB080091, have Enhanced IHC status (HPA antibodies); that supports the observed pattern, while performance of a different antibody still needs its own controls. |
| Isoforms | UniProt lists 2 isoforms and describes isoform 2 as testis-specific (UniProt Q99661 isoforms; tissue specificity). The supplied records do not map antibody epitopes to isoforms, so staining cannot be assigned to either isoform from these data. |
| Mitotic context | KIF2C acts at microtubule plus ends and kinetochores during mitosis (UniProt Q99661 function; subcellular location). Cell-cycle context may help interpret puncta, but those mechanistic locations are not a substitute for the observed HPA tissue IHC pattern. |
| IF/ICC Q: What localisation is reported? | A: Midbody and centrosome are approved; nucleoplasm and cytosol are uncertain (HPA subcellular ICC-IF). These cell-image observations inform IF interpretation but do not establish an IHC tissue pattern or supply an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| The testis positive section has no signal. | The expected High-staining pachytene spermatocytes may be absent from the field (HPA tissue IHC: testis), or an IHC workflow step may have failed (general IHC practice). | Verify cell identity and section quality; then check the antibody's stated IHC conditions, detection reagents and a concurrent positive control (general IHC practice). |
| Only faint signal is seen in an appendix or thymus section. | The named positive populations are reported as Medium, below the testis pachytene reference (HPA tissue IHC: appendix endocrine and thymus cortical cells; testis pachytene spermatocytes). | Locate the named cells and compare with a testis positive section before changing detection conditions (HPA tissue IHC; general IHC practice). |
| A sharp membrane rim dominates. | A surface pattern conflicts with the reported intracellular locations and absence of a transmembrane segment (HPA tissue IHC; UniProt Q99661 topology). | Check staining with a no-primary control and an independently validated IHC antibody where available (general IHC practice; HPA antibodies: HPA079172 and CAB080091 Enhanced for IHC). |
| Strong colour appears in colon endocrine cells. | That population is reported as not detected, so the result needs confirmation; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: colon endocrine cells; general IHC practice). | Check a no-primary control, inspect the cell assignment and compare with the expected testis positive pattern (general IHC practice; HPA tissue IHC: testis). |
| Brown haze obscures cell boundaries. | Diffuse background prevents confident assignment of the HPA-reported compartments and cell populations (HPA tissue IHC; general IHC practice). | Review blocking, washes and detection controls; score localisation only where cell boundaries and nuclei remain interpretable (general IHC practice). |
| Scattered intracellular puncta do not match a simple nuclear or cytoplasmic call. | UniProt reports mitotic kinetochore and microtubule-associated locations, but HPA's tissue summary does not define a punctate IHC scoring pattern (UniProt Q99661 subcellular location; HPA tissue IHC). | Check cell morphology and the expected tissue population; report the observed compartment without assigning puncta to a mitotic structure from chromogenic IHC alone (general IHC practice). |
Experiment evidence
IHC & IF images showing KIF2C staining
PB9230 · IHCAnti-MCAK antibody, PB9230, IHC(P) IHC(P): Rat Thymus Tissue
PB9230 · IHCAnti-MCAK antibody, PB9230, IHC(P) IHC(P): Rat Testis Tissue
PB9230 · IFIF analysis of MCAK using anti-MCAK antibody (PB9230). MCAK was detected in immunocytochemical section of U20S cells. Enzyme antigen retriev
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142945-KIF2C/tissue/cerebral+cortex#imid_21062291
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142945-KIF2C/tissue/colon#imid_21062201
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142945-KIF2C/tissue/liver#imid_21062218
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000142945-KIF2C/tissue/kidney#imid_21062211
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA006219; green = KIF2C, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA006219; green = KIF2C, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KIF2C IHC & IF
🧪Run testis first; pachytene spermatocytes should stain (HPA: High in pachytene spermatocytes). Use adrenal gland glandular cells as a negative tissue (HPA: Not detected); unstained neighboring cells on the testis slide, if present, provide an internal background reference, but other testis cell types should not be presumed negative (HPA: positive call is for pachytene spermatocytes).
Positive control tissue: Testis (Pachytene spermatocytes, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KIF2C in Rh30, SiHa, U2OS, with annotated localisation: Nucleoplasm (uncertain), Midbody (approved), Centrosome (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and immunoglobulin-class-matched isotype controls; confirm specificity with knockout material or peptide competition if a matched peptide is available (standard IHC practice). For testis sections, quench endogenous peroxidase, block endogenous biotin if using avidin–biotin detection, and check autofluorescence for IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the PB9230 rat-thymus IHC(P) caption does not state a fixative (caption: PB9230, rat thymus IHC(P)). Antigen-retrieval dependency is unreported; optimize retrieval using the catalog antibody’s IHC-P protocol (standard IHC practice). Whether frozen sections or IF/ICC are easier is unreported, despite available ICC-IF images in Rh30, SiHa and U2OS (HPA: subcellular images); assess intrinsic fluorescence on testis sections before interpreting an IF signal (standard IF practice).
HPA tissue IHC evidence for KIF2C
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Testis | Pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Thymus | Cortical cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KIF2C IHC Tips
Troubleshoot chromogenic KIF2C staining by checking retrieval, tissue handling, cellular localisation and controls before comparing samples.
What retrieval conditions should I try when KIF2C staining is weak in paraffin sections?
Use heat-mediated EDTA at pH 8.0 for KIF2C antigen retrieval (datasheet PB9230). In a pilot, compare 10 and 20 minutes of heating on adjacent sections, then cool them for 20 minutes while keeping detection conditions constant (standard IHC optimization). If staining remains weak, test citrate at pH 6.0 as a fallback, with the same positive control in each run (standard IHC optimization). Evaluate signal in testis pachytene spermatocytes or thymus cortical cells, which have reported KIF2C staining (HPA tissue IHC). Reject conditions that increase tissue damage or diffuse background more than interpretable cellular signal (standard IHC practice).
Could fixation explain variable KIF2C staining between my paraffin blocks?
Target-specific fixation sensitivity is unknown from the supplied evidence; the rat thymus IHC(P) caption does not state a fixative (PB9230 caption). Record each block's fixative, fixation duration and processing history before comparing staining (standard IHC practice). For a controlled pilot, process matched material with 10% neutral buffered formalin for 6 versus 24 hours, then stain the sections together (standard IHC optimization). Keep retrieval at EDTA pH 8.0 so fixation is the main tested variable (datasheet PB9230; standard IHC optimization). Interpret any difference as a result of that pilot, rather than assuming an established KIF2C-specific fixation effect (standard IHC practice).
Should KIF2C appear in nuclei, cytoplasm or mitotic structures?
Interpret the compartment in the context of tissue and cell state: KIF2C is associated with cytoskeleton, nucleus, centromeres and kinetochores (UniProt Q99661). Tissue IHC reports nuclear expression in testis and cytoplasmic expression in lymphoid tissues and intestines (HPA tissue IHC). Examine mitotic figures separately, because KIF2C regulates kinetochore microtubules during chromosome segregation (UniProt Q99661). Compare nuclear and cytoplasmic staining on adjacent fields at the same chromogen development time, with a hematoxylin counterstain to locate nuclei (standard IHC practice). Diffuse staining across every compartment or cell type warrants a control review before assigning a biological localisation (standard IHC interpretation).
How can an unknown epitope affect interpretation of KIF2C staining?
KIF2C has 2 reported isoforms, and isoform 2 is testis-specific (UniProt Q99661). Without a mapped antibody epitope, staining alone cannot establish which isoform the IHC-validated antibody detects (standard antibody interpretation). The kinesin motor spans residues 258–588, while the record also lists modified residues outside that region (UniProt Q99661). Check the immunogen or epitope documentation, if available, against both isoform sequences before making an isoform claim (standard antibody validation practice). If that mapping remains unavailable, report staining as KIF2C-associated signal and describe the stained cell type and compartment (standard IHC reporting practice).
How should I follow up chromogenic KIF2C IHC with multiplex immunofluorescence?
Use IF/ICC as a separate validation experiment; the catalog evidence here shows rat thymus IHC(P), without specifying a fixative (PB9230 caption). Pair KIF2C with a validated marker for the expected cell type, such as a pachytene spermatocyte marker in testis, and check that both signals occupy appropriate cells (HPA tissue IHC; standard multiplex IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with lower measured tissue autofluorescence (standard multiplex IF practice). Because KIF2C has no transmembrane segment and can occupy nuclear and cytoskeletal sites, optimize permeabilisation for intracellular access (UniProt Q99661; standard IF practice). Include single-stain and secondary-only controls to assess bleed-through and background (standard IF practice).
What should I check when KIF2C chromogenic staining looks diffuse?
Compare the stained section with no-primary and matched detection controls to identify non-specific reagent signal (standard IHC practice). Block endogenous peroxidase before peroxidase-based chromogen development and inspect whether residual colour persists in the no-primary control (standard chromogenic IHC practice). Titrate the primary antibody and shorten DAB development if colour fills extracellular space or obscures cell boundaries (standard IHC optimization). Check for drying, folds and edge-heavy staining on the section before treating those patterns as KIF2C localisation (standard IHC interpretation). Use a reported positive population, such as thymus cortical cells, to preserve a meaningful signal while reducing background (HPA tissue IHC).
How should I quantify KIF2C staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment and cell population before counting; testis nuclear and lymphoid cytoplasmic patterns differ in the tissue evidence (HPA tissue IHC). For a defined cell population, report the percentage of positive cells and an H-score using 0–3 intensity categories, with an H-score range of 0–300 (standard IHC scoring practice). For sparse mitotic structures, count positive cells per mm² of viable tissue alongside the total cells assessed (standard IHC quantification practice). Normalize comparisons to the same eligible cell type and viable area, rather than whole-section area when cellular composition varies (standard IHC practice). Hold retrieval, exposure to chromogen and scoring thresholds constant across the comparison (standard IHC quantification practice).
How can I distinguish true KIF2C staining from artefact?
Prioritize staining in reported populations, including testis pachytene spermatocytes and thymus cortical cells, when assessing plausibility (HPA tissue IHC). Consider the compartment too: testis nuclear and lymphoid cytoplasmic patterns are reported, and KIF2C can associate with mitotic centromeres and kinetochores (HPA tissue IHC; UniProt Q99661). Compare suspicious staining with a no-primary control, particularly where endogenous peroxidase could produce chromogen without primary antibody (standard chromogenic IHC practice). Exclude section edges, folds and necrotic areas from scoring when they show irregular, cell-independent colour (standard IHC interpretation). Treat signal confined to an unexpected cell type or compartment as unconfirmed until supported by appropriate controls (standard IHC interpretation).
Boster reagents
Best KIF2C / Kinesin-like protein KIF2C IHC Antibodies
PB9230 has paraffin-section IHC images from rat thymus and rat, mouse, and human testis (catalog IHC captions), plus an IF image from U20S cells (catalog IF caption).
Real IHC data
Anti-MCAK/KIF2C Antibody ®
Cat # PB9230
The only card that will render is PB9230, which is listed for IHC and IF in human, mouse, and rat (catalog applications and reactivity). Its figures show paraffin-section IHC in rat thymus and rat, mouse, and human testis, and IF in U20S cells (catalog image captions).
Which to pick: Choose PB9230 for tissue IHC: its own captions document paraffin sections from human, mouse, and rat, but do not report the fixative (catalog IHC captions). For IF/ICC, PB9230 has a U20S IF image; M03202-1 is a human-reactive rabbit monoclonal listed for IF/ICC but has no image in this payload (catalog IF captions, applications, reactivity, and clone). PB9230 is the cross-species choice because its listed reactivity spans human, mouse, and rat (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99661 (KIF2C_HUMAN, Kinesin-like protein KIF2C).
- Human Protein Atlas. KIF2C tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. KIF2C subcellular location (ICC-IF): Mainly localized to the nucleoplasm, centrosome and midbody. In addition localized to the cytosol..
- Human Protein Atlas. KIF2C antibody validation summary (3 antibodies).
- Bioinformatical analysis of the key differentially expressed genes for screening potential biomarkers in Wilms tumor. Scientific reports 2023 — PMC10505208.
- Characterization of Kinesin Family Member 2C as a Proto-Oncogene in Cervical Cancer. Frontiers in pharmacology 2021 — PMC8828917.
- KIF2C Facilitates Tumor Growth and Metastasis in Pancreatic Ductal Adenocarcinoma. Cancers 2023 — PMC10000478.
- KIF2C is a Biomarker Correlated With Prognosis and Immunosuppressive Microenvironment in Human Tumors. Frontiers in genetics 2022 — PMC9171145.
- PubMed PMID:9434124 — UniProt-cited evidence.
- PubMed PMID:12383881 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KIF2C (Kinesin-like protein KIF2C). Expected localisation Nuclear in testis; cytoplasmic in lymphoid tissues and intestines (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9230). Validated IHC antibody: PB9230 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Pachytene spermatocytes); negative Adrenal gland. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PB9230); 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 PB9230); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9230); 8. Primary antibody: Rabbit anti-KIF2C, 0.5-1μg/ml (datasheet PB9230); 9. Primary incubation: Overnight at 4 °C (datasheet PB9230). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9230); Caveat: Staining intensity and compartment vary by tissue (HPA tissue IHC); Isoform / epitope: Two isoforms; isoform 2 is testis-specific. Check epitope coverage (UniProt); Regulation: Highest staining in testis; lower in other sampled tissues (HPA tissue IHC). Evidence: UniProt Q99661, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm, centrosome and midbody. In…, Datasheet PB9230, PMC10505208, PMC8828917. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KIF2C/immunohistochemistry-ihc-guide.