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KANK2 / KN motif and ankyrin repeat domain-containing protein 2 · IHC design guide
Design Immunohistochemistry for KANK2
Plan chromogenic IHC for KANK2 using its observed cytoplasmic tissue pattern and cell-specific controls (HPA tissue IHC). Compare staining in cervical squamous epithelial cells, where expression is high, with pancreatic exocrine glandular cells, where it is undetected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KANK2 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in squamous epithelia and smooth muscle (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | Pancreas+2 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) | |
| Caveat | Staining shows low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Pro-apoptotic stimuli promote degradation (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended KANK2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published KANK2 staining methods for tumor and mouse lung sections (PMC11786046; PMC8946178).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07999-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-KANK2, 1:100–1:500 starting range (standard) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KANK2-positive staining in squamous epithelial cells of cervix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, including squamous epithelia and smooth muscle. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the published excerpts give no retrieval conditions (PMC11786046; PMC8946178).
Section 2
What Is the Expected KANK2 Staining Pattern?
For paraffin-section IHC, expect mainly cytoplasmic KANK2 staining, especially in cervical and esophageal squamous epithelial cells and endometrial stromal cells (HPA tissue IHC: High; cytoplasmic profile). KANK2 has no transmembrane segment, and UniProt lists cytoplasmic and mitochondrial locations (UniProt Q63ZY3: topology; subcellular location). Interpret the tissue pattern cautiously: HPA rates its IHC evidence Approved but reports low consistency between antibody staining and RNA expression, pending external verification (HPA tissue IHC: reliability).
| Distinct cytoplasmic staining in cervical or esophageal squamous epithelium, or endometrial stromal cells. | This fits the reported high IHC signal in those cell populations and the tissue-wide cytoplasmic profile (HPA tissue IHC: High; cytoplasmic profile). Judge the named cells, rather than assigning one intensity to every cell in the section. |
| Predominantly nuclear staining replaces the expected cytoplasmic pattern in a positive IHC tissue. | Treat this as an IHC localisation mismatch and check morphology, detection background and antibody specificity (HPA tissue IHC: cytoplasmic profile). Nuclear signal alone is not proof of artefact: HPA reports approved nucleoplasmic localisation in ICC-IF (HPA subcellular: Nucleoplasm approved). |
| Strong signal appears in a cell population reported as unstained, such as pancreatic exocrine glandular cells. | Investigate antibody cross-reactivity or endogenous chromogenic detection activity before calling it KANK2 (HPA tissue IHC: pancreas, Not detected; standard IHC practice). A tissue-level result does not validate every unexpected cell-level signal. |
| Colour spreads across the section without clear cell boundaries or a reproducible intracellular pattern. | This is background rather than a convincing KANK2 pattern; inspect the no-primary control and review blocking, washes and detection conditions (standard IHC practice). HPA's reported pattern is cellular and cytoplasmic (HPA tissue IHC: cytoplasmic profile). |
| No staining appears in cervical or esophageal squamous cells when the run is otherwise readable. | A failed positive control is possible because both populations are reported High (HPA tissue IHC: cervix and esophagus). Review tissue identity, antibody application and the run controls before treating the test section as KANK2-negative (standard IHC practice). |
💡Expected KANK2 appearanceCall an IHC result consistent when identifiable cervical or esophageal squamous cells, or endometrial stromal cells, show clear cytoplasmic staining with strong signal relative to local background (HPA tissue IHC: High; cytoplasmic profile); widespread cell-free colour or dominant nuclear-only IHC staining warrants investigation (standard IHC practice; HPA tissue IHC: cytoplasmic profile).
| Which compartment should I use for IHC scoring? | Use the cytoplasmic tissue-IHC profile for paraffin sections (HPA tissue IHC: cytoplasmic profile). UniProt also lists mitochondria, but the supplied tissue-IHC record does not resolve a mitochondrial pattern (UniProt Q63ZY3: subcellular location; HPA tissue IHC: profile). |
| What if I compare the separate IF/ICC guide? | HPA ICC-IF reports mainly nucleoplasm, with additional cytosol (HPA subcellular: Nucleoplasm approved; Cytosol supported). That assay-specific observation merits comparison, not automatic transfer into an IHC scoring rule; HPA015643 is IHC Approved and ICC Uncertain (HPA antibodies). |
| How firm is tissue selection? | High staining is reported in three named cell populations, while pancreatic exocrine, stomach glandular and bladder urothelial cells are Not detected (HPA tissue IHC). HPA also reports low tissue RNA specificity and low staining–RNA consistency, so these are comparison tissues, not absolute biological controls (HPA tissue IHC: reliability; RNA specificity). |
| Do protein features predict a different slide pattern? | UniProt records three isoforms, no signal peptide or propeptide, and no transmembrane segment (UniProt Q63ZY3: isoforms; processing; topology). Those annotations do not establish which isoform an antibody detects or predict fixation sensitivity; the supplied evidence gives neither an epitope map nor a target-specific fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive comparison tissue is blank. | The expected High cell population may be absent from the examined area, or the IHC run may have failed (HPA tissue IHC: cervix, esophagus and endometrium High; standard IHC practice). | Confirm the named cells on the counterstain, then check the antibody and detection controls and repeat alongside a known-positive section (standard IHC practice). |
| Colour is diffuse or covers tissue and empty spaces. | Nonspecific binding or detection background can obscure the reported cytoplasmic pattern (standard IHC practice; HPA tissue IHC: cytoplasmic profile). | Compare a no-primary control; review blocking, washing and detection exposure, then score only discrete cellular signal (standard IHC practice). |
| Nuclei dominate the paraffin-section stain. | The result differs from HPA tissue IHC, although ICC-IF separately reports nucleoplasmic KANK2 (HPA tissue IHC: cytoplasmic profile; HPA subcellular: Nucleoplasm approved). | Check the no-primary control and repeat IHC in a reported high-staining tissue before interpreting the compartment (standard IHC practice; HPA tissue IHC: High). |
| Pancreatic exocrine cells stain strongly. | This conflicts with their Not detected HPA IHC result; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: pancreas; standard IHC practice). | Check detection-only and no-primary controls, then compare staining in a reported high-staining cell population (standard IHC practice; HPA tissue IHC: High). |
| A low-staining tissue seems negative. | A faint signal may be hard to separate from background in cells HPA rates Low (HPA tissue IHC: cerebellar granular-layer cells, caudate glia and others Low). | Use a reported High cell population to assess run performance; avoid treating a Low tissue alone as a failed assay (HPA tissue IHC: High and Low; standard IHC practice). |
| Staining varies between sections despite readable controls. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Compare matched cell populations and run controls, record the variation, and avoid inferring a target-specific fixation effect from these data (standard IHC practice; HPA tissue IHC: reliability). |
Experiment evidence
IHC & IF images showing KANK2 staining
A07999-1 · IHCImmunohistochemistry of KANK2 in human kidney tissue with KANK2 antibody at 2.5 μg/ml.
A07999-1 · IFImmunofluorescence of KANK2 in human kidney tissue with KANK2 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000197256-KANK2/tissue/cerebral+cortex#imid_4029020
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000197256-KANK2/tissue/colon#imid_4028946
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000197256-KANK2/tissue/liver#imid_4028955
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000197256-KANK2/tissue/kidney#imid_4028948
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA015643; green = KANK2, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA015643; green = KANK2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KANK2 IHC & IF
🧪Run cervix first and require staining in its squamous epithelial cells (HPA: High). Run pancreas as the negative tissue, focusing on exocrine glandular cells (HPA: Not detected); on the cervix slide, treat cells outside the positive squamous epithelium as internal background checks, without assuming they are validated KANK2-negative cells (HPA: High in cervical squamous epithelial cells).
Positive control tissue: Cervix (Squamous epithelial cells, HPA High)
Negative control tissue: Pancreas (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KANK2 in Rh30, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a host- and immunoglobulin-matched isotype control; use KANK2-knockout tissue or cells as a biological negative where available (standard IHC practice). Quench endogenous peroxidase and inspect blood-rich areas of the cervix slide for chromogenic background (standard IHC practice).
⚠️Feasibility: No KANK2-specific fixation window or antigen-retrieval effect is reported in the supplied evidence; the selected A07999-1 kidney IHC caption does not state its fixative (selected tissue-IHC caption). Optimize retrieval empirically for paraffin sections; the supplied evidence does not establish that frozen sections or IF/ICC are easier (selected tissue-IHC caption; HPA subcellular: ICC-IF images available). In cervix, blood-associated endogenous peroxidase can resemble positive chromogen, so interpret staining in the squamous epithelial cells against the controls (standard IHC practice; HPA: High in cervical squamous epithelial cells).
HPA tissue IHC evidence for KANK2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced KANK2 IHC Tips
Troubleshoot KANK2 chromogenic IHC in paraffin sections by checking retrieval, compartment, cell identity and staining controls before interpreting intensity.
How should I retrieve KANK2 when paraffin sections stain weakly?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Let sections cool in the retrieval buffer, then compare the same tissue and antibody concentration across runs to distinguish retrieval failure from detection variability (standard IHC practice). The selected kidney image reports 2.5 µg/ml antibody but gives no retrieval conditions, so it cannot verify this heating step (A07999-1 caption). If signal remains weak, test a more alkaline retrieval buffer on adjacent sections as a fallback while watching for tissue damage and increased background (standard IHC practice).
Could fixation explain weak or uneven KANK2 staining?
KANK2-specific fixation sensitivity is unknown: the selected kidney caption does not state a fixative, and the supplied evidence does not establish a target-specific fixation effect (A07999-1 caption). For paraffin IHC, record the fixative, fixation duration and processing history before comparing specimens, because these variables can change antigen accessibility (standard IHC practice). Compare adjacent sections from the same block under identical citrate pH 6.0 retrieval and detection conditions when investigating uneven staining (page retrieval setting; standard IHC practice). If fixation histories differ, describe that limitation in the result rather than attributing a signal difference to KANK2 abundance (standard IHC practice).
Which KANK2 staining compartment should count as plausible in tissue?
Prioritise interpretable cytoplasmic staining in morphologically intact cells: tissue IHC describes cytoplasmic expression, while UniProt also lists mitochondrial localisation (HPA tissue IHC; UniProt Q63ZY3 localisation). KANK2 has no annotated transmembrane segment, so a crisp membrane-only rim needs independent validation before it is scored as target signal (UniProt Q63ZY3 topology; standard IHC practice). Subcellular IF data instead report nucleoplasm as the main location and cytosol as an additional location, making isolated nuclear IHC staining ambiguous (HPA subcellular). Check nuclear counterstain and cell borders on matched sections, and document cytoplasmic and nuclear scores separately if both persist (standard IHC practice).
How do KANK2 isoforms and epitope placement affect IHC interpretation?
KANK2 has 3 listed isoforms, but the supplied antibody caption does not identify the recognised epitope or its isoform coverage (UniProt Q63ZY3 isoforms; A07999-1 caption). Obtain an epitope map for the catalog antibody before treating staining as a measure of every isoform (standard IHC practice). The protein has no annotated transmembrane segment and has reported phosphorylation sites, including serines 19 and 83; those facts alone do not establish epitope sensitivity (UniProt Q63ZY3 topology and modified residues). If results conflict across specimens, compare independently validated antibodies against documented regions on serial sections and report which reagent produced each score (standard IHC practice).
How can IF help check an uncertain KANK2 IHC pattern?
Use IF as an orthogonal localisation check when chromogenic IHC leaves cell identity or compartment uncertain (standard IHC/IF practice). Multiplex KANK2 with a validated marker for the expected cell population; for a kidney question, distinguish podocytes from mesangial cells because both express KANK2 at protein level (UniProt Q63ZY3 tissue specificity). Select fluorophores outside the strongest measured tissue autofluorescence and include single-colour controls to assess bleed-through (standard IF practice). KANK2 lacks a transmembrane segment, so permit intracellular antibody access with a compatible permeabilisation step, then assess cytosolic, mitochondrial and nuclear patterns without assuming the IHC antibody has been validated for IF (UniProt Q63ZY3 topology and localisation; HPA subcellular; standard IF practice).
What should I check when KANK2 IHC shows diffuse background?
Run a no-primary control and inspect whether colour persists in the same compartments; residual colour points toward detection chemistry or tissue background (standard IHC practice). Apply a peroxidase block before chromogenic detection, then check whether excessive DAB development, insufficient washing or overly concentrated primary antibody explains diffuse colour (standard IHC practice). The selected kidney image used 2.5 µg/ml, which is an image condition rather than a universal working concentration (A07999-1 caption). Compare intact cell cytoplasm with tissue edges and damaged areas, and adjust one variable at a time while preserving the citrate pH 6.0 retrieval baseline (HPA tissue IHC; page retrieval setting; standard IHC practice).
How should I score KANK2 across heterogeneous IHC sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; cytoplasmic tissue staining is reported, and kidney podocytes and mesangial cells both have protein-level expression evidence (HPA tissue IHC; UniProt Q63ZY3 tissue specificity). An H-score combines the percentages of cells at each intensity, whereas percent-positive cells or positive-cell density per mm² can answer different questions (standard IHC practice). Normalise each measure to the number of eligible cells or the analysed tissue area, and use the same threshold and counterstain criteria across sections (standard IHC practice). Report compartments separately and retain representative images because tissue staining and RNA data have low consistency in the supplied assessment (HPA tissue IHC reliability; standard IHC practice).
When is apparent KANK2 positivity likely to be an artefact?
A convincing result has signal in intact, identifiable cells with a reproducible compartment pattern; tissue IHC reports cytoplasmic expression in several tissues (HPA tissue IHC; standard IHC practice). Strong staining confined to a membrane rim is questionable for a protein with no annotated transmembrane segment, while nuclear-only staining needs caution given differing tissue IHC and IF localisation reports (UniProt Q63ZY3 topology; HPA tissue IHC; HPA subcellular). Exclude section-edge enhancement, necrotic regions and colour that survives the no-primary control, including possible endogenous enzyme signal (standard IHC practice). Compare expected positive cell populations with less reactive populations and avoid claiming specificity solely from intensity because antibody staining and RNA expression show low consistency (HPA tissue IHC reliability; standard IHC practice).
Boster reagents
Best KANK2 / KN motif and ankyrin repeat domain-containing protein 2 IHC Antibodies
A07999-1 has IHC-P and IF applications (catalog: applications), with images from human kidney tissue (catalog: IHC and IF image captions). Human, mouse, and rat are listed as reactive (catalog: reactivity).
Real IHC data
Anti-KANK2 Antibody
Cat # A07999-1
A07999-1 will render with an IHC image of human kidney tissue stained at 2.5 μg/ml (catalog: IHC image caption). Its IF image also shows human kidney tissue, stained at 20 μg/ml (catalog: IF image caption).
Which to pick: For paraffin-section tissue IHC, choose A07999-1 (catalog: IHC-P application); its human kidney IHC caption does not report the fixative (catalog: IHC image caption). For IF, A07999-1 has a human kidney image (catalog: IF image caption); ICC validation is unreported (catalog: applications and image captions). Human, mouse, and rat reactivity is listed for A07999-1, but its IHC and IF images document human tissue only (catalog: reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q63ZY3 (KANK2_HUMAN, KN motif and ankyrin repeat domain-containing protein 2).
- Human Protein Atlas. KANK2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KANK2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. KANK2 antibody validation summary (1 antibodies).
- Pan-Cancer Analysis of KANK2: Clinical and Molecular Insights into Tumor Progression and Therapeutic Implications. Journal of Cancer 2025 — PMC11786046.
- HSP70 Ameliorates Septic Lung Injury via Inhibition of Apoptosis by Interacting with KANK2. Biomolecules 2022 — PMC8946178.
- PubMed PMID:17476305 — UniProt-cited evidence.
- PubMed PMID:10819331 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KANK2 (KN motif and ankyrin repeat domain-containing protein 2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07999-1 at 1:100–1:500 (standard starting range) (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Squamous epithelial cells); negative Pancreas. 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 A07999-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-KANK2, 1:100–1:500 starting range (standard); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining shows low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms ; epitope coverage is unknown (UniProt); Regulation: Pro-apoptotic stimuli promote degradation (UniProt). Evidence: UniProt Q63ZY3, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A07999-1, PMC11786046, PMC8946178. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KANK2/immunohistochemistry-ihc-guide.