KIF2C / Kinesin-like protein KIF2C · IHC design guide

Design Immunohistochemistry for KIF2C

Plan KIF2C staining in paraffin sections using the expected nuclear testis and cytoplasmic lymphoid patterns (HPA tissue IHC). This guide covers tissue controls, fixation consistency, detection, and interpretation of low-level staining.

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for KIF2C (IHC for KIF2C): expected localisation Nuclear in testis; cytoplasmic in lymphoid tissues and intestines (HPA tissue IHC), antibody PB9230, validated IHC image, and IHC protocol steps — Printable **KIF2C** IHC protocol sheet — expected localisation Nuclear in testis; cytoplasmic in lymphoid tissues and intestines (HPA tissue IHC), antibody PB9230, controls and protocol steps. Open the full KIF2C IHC guide →

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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	Pachytene spermatocyte nuclei; lymphoid cytoplasm (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	Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9230)
	Caveat	Low germinal-center staining may be missed (HPA tissue IHC)
	Regulation	High in testis and thymus; low in gut (UniProt)
	Isoform / epitope	2 isoforms; isoform 2 is testis-specific; epitope coverage is unknown (UniProt)

Section 1

Recommended KIF2C IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by published KIF2C IHC methods for Wilms tumor, tissue sections, and liver cancer (PMC10505208; PMC8828917; PMC9171145).

Recommended immunohistochemistry (IHC-P) protocol parameters
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 retrieval at pH 8.0 (PB9230 datasheet); use citrate retrieval when following the published Wilms tumor or tissue-section methods (PMC10505208; PMC8828917).

Section 2

What Is the Expected KIF2C Staining Pattern?

KIF2C is a nonmembrane protein associated with the cytoskeleton, nucleus, centromere and kinetochore (UniProt Q99661: localization and topology). In IHC, expect strong nuclear staining in testicular pachytene spermatocytes and moderate staining in thymic cortical cells; HPA also describes cytoplasmic expression in lymphoid tissues and intestines (HPA: tissue IHC, Enhanced reliability). Interpret the stained cell type alongside its compartment, because the reported pattern varies by tissue (HPA: tissue IHC).

What am I looking at on my slide?
Strong nuclear staining in pachytene spermatocytes, with weaker or absent staining in surrounding cells (HPA: testis, High).	This is the clearest supplied positive tissue pattern. Evaluate the named cells, rather than requiring uniform staining across the section (HPA: testis, High; general IHC practice).
Moderate staining in thymic cortical cells or appendix endocrine cells (HPA: thymus and appendix, Medium).	These are additional observed positives. Cytoplasmic signal can be compatible with the HPA lymphoid tissue profile; a testis-like nuclear pattern is not required in every tissue (HPA: tissue IHC profile).
Predominantly membrane-outline or extracellular staining, without staining in an expected cellular compartment.	Treat this as a possible artefact and review controls: KIF2C has no transmembrane segment or signal peptide, and its reported locations are intracellular (UniProt Q99661: topology, processing and localization).
Strong staining in cells listed as undetected, or broad colour deposition across unrelated cell types (HPA: negative tissue IHC entries).	Cross-reactivity or endogenous detection activity is possible, not proven. Check the exact cell type and a no-primary control before assigning the signal to KIF2C (HPA: tissue IHC; general IHC practice).
No signal in pachytene spermatocytes in a testis section (HPA: testis, High).	The run lacks its strongest supplied positive reference. Check tissue preservation, retrieval, primary antibody conditions and detection controls before calling an unknown sample negative (general IHC practice).

💡Expected KIF2C appearanceA convincing positive shows High nuclear staining in testicular pachytene spermatocytes (HPA: testis), while membrane-outline or extracellular staining without intracellular signal warrants artefact checks (UniProt Q99661: topology and localization; general IHC practice).

How each factor affects the staining
Tissue and cell selection (HPA: tissue IHC).	Testicular pachytene spermatocytes are High; thymic cortical and appendix endocrine cells are Medium. Bone marrow hematopoietic cells are Low, so a weak result there is less decisive (HPA: tissue IHC).
IHC antibody evidence (HPA: antibody validation).	HPA079172 and CAB080091 have Enhanced IHC validation; HPA006219 has Approved ICC status but no IHC status in the supplied record. Match the antibody's validation to the assay (HPA: antibody validation; general IHC practice).
Intracellular target and processing (UniProt Q99661: topology and processing).	KIF2C has no transmembrane segment or signal peptide; the reported chain spans residues 2–725. These facts support intracellular interpretation but do not identify the antibody epitope or predict antigen retrieval response (UniProt Q99661; general IHC practice).
Isoform distribution (UniProt Q99661: isoforms and tissue specificity).	UniProt lists two isoforms and describes isoform 2 as testis-specific. The supplied evidence does not show which isoforms the IHC antibodies recognize, so testis staining cannot be assigned to one isoform (UniProt Q99661; HPA: antibody validation).
IF/ICC Q&A: What localization is reported?	HPA reports approved centrosome and midbody localization; nucleoplasm and cytosol are uncertain. Its ICC images include Rh30, SiHa and U2OS. These IF observations do not define an IHC-P protocol (HPA: subcellular ICC-IF).
Antigen retrieval (general IHC practice).	Follow the IHC-validated antibody's stated retrieval conditions and assess them with a positive control. The supplied UniProt and HPA records give no KIF2C-specific fixation or retrieval sensitivity (general IHC practice; UniProt Q99661; HPA: tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Testis positive control is blank (HPA: pachytene spermatocytes, High).	A failed staining run or unsuitable antibody conditions are possible; the slide alone cannot identify which step failed (general IHC practice).	Check control tissue, the IHC-validated antibody's retrieval and dilution instructions, and detection reagents; repeat with a known-positive section (general IHC practice).
Weak signal in bone marrow hematopoietic cells (HPA: bone marrow, Low).	Low expression is compatible with the HPA observation; intensity alone does not establish an assay failure (HPA: tissue IHC).	Compare with testicular pachytene spermatocytes on the same run before interpreting a weak unknown sample (HPA: testis, High; general IHC practice).
Diffuse chromogen obscures cellular boundaries.	Nonspecific antibody binding, endogenous enzyme activity or excess detection signal may contribute (general IHC practice).	Review no-primary controls, blocking and detection steps; adjust antibody conditions according to the IHC-validated antibody's instructions (general IHC practice).
Strong staining appears in an HPA-undetected cell type (HPA: tissue IHC negative entries).	Cross-reactivity or endogenous detection activity is possible; an HPA negative entry alone cannot diagnose the mechanism (HPA: tissue IHC; general IHC practice).	Confirm the cell identity, inspect a no-primary control and compare an independent IHC-validated antibody where available (HPA: antibody validation; general IHC practice).
Signal outlines membranes or extracellular material while expected cells remain unstained.	This conflicts with the reported intracellular locations and lack of a transmembrane segment or signal peptide (UniProt Q99661: localization, topology and processing).	Check controls and chromogen deposition, then reassess the compartment in a known-positive testis section (HPA: testis, High; general IHC practice).
IHC appears to disagree with ICC-IF localization.	HPA reports nuclear testis and cytoplasmic lymphoid or intestinal tissue patterns; its ICC centrosome and midbody findings come from a different assay (HPA: tissue IHC and subcellular ICC-IF).	Interpret each result against its own tissue, cell type and assay validation; use IHC-validated antibodies for the paraffin section comparison (HPA: antibody validation; general IHC practice).

Source: UniProt Q99661 Human Protein Atlas tissue Human Protein Atlas subcellular

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

IF analysis of MCAK using anti-MCAK antibody (PB9230). MCAK was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-MCAK Antibody (PB9230) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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.

← 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 strongly (HPA: High in testis pachytene spermatocytes). Use adrenal gland as the negative tissue (HPA: glandular cells Not detected); on the testis slide, assess non-pachytene cells for background staining without assuming every such cell is KIF2C-negative (HPA: positive call is specific to 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 a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, clonality and concentration; and KIF2C-knockout material or an immunizing-peptide block if the peptide is available (standard IHC practice). In testis, quench endogenous peroxidase and check for endogenous biotin background if using biotin-based detection (standard chromogenic IHC practice).

⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval for the IHC-validated antibody (supplied target/application evidence; standard IHC practice). The PB9230 rat-thymus IHC(P) caption does not report a fixative, and the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (selected-SKU caption; supplied target/application evidence). Dense germ-cell layers can complicate assignment of staining to individual testis cells, so use a nuclear counterstain and tissue morphology when scoring (standard IHC 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 KIF2C staining in paraffin sections by checking retrieval, cellular compartment, tissue context and controls before interpreting chromogenic signal.

How should I troubleshoot weak KIF2C staining after antigen retrieval?

Start with heat-mediated EDTA retrieval at pH 8.0 for this paraffin-section assay (datasheet PB9230). If staining is weak, compare a modest range of heating and cooling times on adjacent sections while keeping the buffer and detection conditions fixed (standard IHC practice). Include a known positive section, such as thymus or testis, in each comparison (HPA tissue IHC: thymus cortical cells Medium; testis pachytene spermatocytes High). Score both signal and tissue damage, since harsh retrieval can impair morphology and increase nonspecific staining (standard IHC practice). Record the final heating time, temperature and cooling interval so the chosen condition can be reproduced (standard IHC practice).

Could fixation explain a weak or uneven KIF2C result?

The supplied evidence does not establish KIF2C-specific fixation sensitivity, and the PB9230 rat-thymus paraffin-section caption does not state a fixative (PB9230 tissue-IHC caption). Document the fixative and fixation duration for each specimen before comparing staining across batches (standard IHC practice). Check whether weak areas coincide with visibly poor preservation, thick sections or uneven processing, using a morphology stain and adjacent sections where available (standard IHC practice). Compare specimens with similar processing histories under the same pH 8.0 EDTA retrieval condition (datasheet PB9230; standard IHC practice). Treat a processing association as a troubleshooting clue rather than proof of a KIF2C-specific fixation effect (standard IHC practice).

Which cellular compartments should contain convincing KIF2C staining?

Interpret chromogenic staining at the cell level: KIF2C is associated with cytoplasm, cytoskeleton, nucleus and mitotic centromeres or kinetochores (UniProt Q99661 subcellular location). In tissue IHC, reported expression is nuclear in testis and cytoplasmic in lymphoid tissues and intestines, so a single compartment rule would be misleading (HPA tissue IHC profile). Examine stained cells against mitotic figures and surrounding architecture before calling puncta kinetochore signal (UniProt Q99661 subcellular location; standard IHC practice). HPA also reports centrosome and midbody localisation in cell imaging, but those fine structures may be difficult to resolve with chromogenic tissue IHC (HPA subcellular; standard IHC practice).

Could isoforms or epitope accessibility alter the KIF2C IHC pattern?

KIF2C has 2 reported isoforms, and isoform 2 is testis-specific (UniProt Q99661 isoforms and tissue specificity). Its kinesin motor domain spans residues 258–588, while multiple modified residues are reported elsewhere in the protein (UniProt Q99661 domains and modified residues). The supplied antibody evidence does not identify the recognized epitope, so it cannot establish which isoforms or modified forms PB9230 detects (PB9230 evidence supplied). If testis and thymus differ, compare staining with the expected cell populations before attributing the difference to isoform recognition (HPA tissue IHC; standard IHC practice). Seek antibody epitope mapping or an independent antibody before making an isoform-specific IHC claim (standard IHC practice).

How should I check KIF2C localisation in a complementary IF experiment?

Use IF as a complementary localisation experiment and distinguish its results from PB9230 paraffin-section IHC evidence (PB9230 tissue-IHC caption; standard IF practice). Multiplex KIF2C with a validated marker identifying the expected cell population, then inspect individual channels before interpreting overlap (HPA tissue IHC: testis pachytene spermatocytes High; standard IF practice). Choose fluorophores after measuring the specimen’s autofluorescence, placing the weaker target signal in a relatively clear channel (standard IF practice). Because KIF2C has no transmembrane segment and has intracellular locations, optimize permeabilisation for access to its cytoplasmic and nuclear epitopes (UniProt Q99661 topology and subcellular location; standard IF practice). Include single-label controls to assess bleed-through and background (standard IF practice).

How can I separate KIF2C signal from chromogenic background?

Run a no-primary control through the same secondary reagent and chromogen steps to locate detection-system background (standard IHC practice). For peroxidase-based detection, apply a peroxidase block and check whether staining remains in the no-primary section (standard IHC practice). Compare background at section edges, folds and damaged areas with intact tissue before scoring cells (standard IHC practice). Titrate the primary antibody and adjust blocking or wash conditions if diffuse staining persists, recording each change separately (standard IHC practice). Use the reported KIF2C-positive cell populations as an anatomical check, while remembering that a plausible location alone does not prove antibody specificity (HPA tissue IHC; standard IHC practice).

What should I measure when comparing KIF2C across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the scored cell population and compartment before reviewing treatment groups, because reported KIF2C patterns differ between testis and lymphoid tissue (HPA tissue IHC profile; standard IHC practice). For chromogenic sections, report the percentage of positive cells and an H-score based on staining intensity, or count positive cells per mm² when cell density is the outcome (standard IHC practice). Normalize counts to the area or number of evaluable cells in the same annotated compartment (standard IHC practice). Exclude folds, necrosis and section edges using the same rules for every specimen (standard IHC practice). Keep retrieval, imaging and scoring thresholds consistent across the comparison (standard IHC practice).

When is a KIF2C-positive IHC result likely to be artefactual?

Look for signal in biologically plausible cells and compartments: HPA reports high staining in testis pachytene spermatocytes and medium staining in thymus cortical cells (HPA tissue IHC). KIF2C can occupy nuclear, cytoskeletal and mitotic structures, so compartment alone cannot validate every stained cell (UniProt Q99661 subcellular location). Uniform staining of cell types reported as negative, such as adrenal glandular cells, warrants scrutiny rather than automatic interpretation as KIF2C (HPA tissue IHC: adrenal glandular cells Not detected; standard IHC practice). Check section edges and necrotic regions for nonspecific deposits, and compare a no-primary control for endogenous enzyme or detection background (standard IHC practice). Require reproducible cellular staining across well-preserved sections before scoring a disputed pattern (standard IHC practice).

Boster reagents

Best KIF2C / Kinesin-like protein KIF2C IHC Antibodies

PB9230 has paraffin-section IHC images from rat, mouse, and human tissues and an IF image from U20S cells (PB9230 IHC/IF captions).

Real IHC data Anti-MCAK antibody, PB9230, IHC(P) IHC(P): Rat Thymus Tissue

Anti-MCAK/KIF2C Antibody ®

Cat # PB9230

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PB9230 is the only card that will render (cards: PB9230); its IHC images show rat thymus, rat testis, mouse testis, and human testis (PB9230 IHC captions). It lists IF/ICC applications and human, mouse, and rat reactivity (PB9230 catalog: applications/reactivity); its IF image shows U20S cells (PB9230 IF caption).

Which to pick: Choose PB9230 for tissue IHC: its captions show paraffin sections, but do not report the fixative (PB9230 IHC captions). For IF/ICC, PB9230 has an IF image of U20S cells (PB9230 IF caption); M03202-1 lists human ICC/IF but has no figure in the payload (M03202-1 catalog: applications/reactivity/image alts). For cross-species work, PB9230 lists human, mouse, and rat reactivity, while M03202-1 lists human reactivity only (PB9230 and M03202-1 catalogs: 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.