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KPNA3 / Importin subunit alpha-4 · IHC design guide
Design Immunohistochemistry for KPNA3
Plan KPNA3 paraffin IHC around nuclear and cytoplasmic staining in most tissues (HPA tissue IHC). Heart cardiomyocytes offer a high-staining reference (HPA tissue IHC); the IHC-validated antibody has a suggested dilution of 1:50–1:100 (datasheet A06131-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KPNA3 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 and cytoplasmic in most tissues (HPA tissue IHC) | |
| Staining pattern | Most tissues show nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A06131-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 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 A06131-1) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Tissue enhanced in skeletal muscle and tongue (HPA tissue RNA) | |
| Isoform / epitope | No isoforms listed; mature chain spans residues 2–521 (UniProt) |
Section 1
Recommended KPNA3 IHC & IF Protocols
The catalog antibody has a datasheet IHC-P protocol (datasheet A06131-1). Three published KPNA3 IHC methods provide additional examples (PMC10431729; PMC6692625; PMC7876302).
| Sample | Paraffin-embedded rat stomach tissue; fixative not specified (datasheet A06131-1) |
| Fixation | Image fixative and duration unreported (datasheet A06131-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A06131-1); 20 min, 95–100 °C (standard) |
| 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-KPNA3, 1:50-1:100 (datasheet A06131-1) |
| 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 | KPNA3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6.0 for the catalog antibody (datasheet A06131-1); assess staining with tissue controls.
Section 2
What Is the Expected KPNA3 Staining Pattern?
KPNA3 staining in paraffin sections is expected in nuclei and cytoplasm across most tissues, with cell-dependent intensity (HPA tissue IHC: Approved; medium consistency with RNA). Nuclear and cytoplasmic localization is consistent with its protein-import role (UniProt O00505: function and subcellular location). KPNA3 has no transmembrane segment, so a membrane-only pattern is unexpected (UniProt O00505: topology).
| Nuclear staining with some cytoplasmic staining in the expected cells. | This fits the reported IHC pattern in most tissues (HPA tissue IHC: nuclear and cytoplasmic expression). Judge intensity within the identified cell population: HPA reports High staining in heart cardiomyocytes and pancreatic exocrine glandular cells, among others (HPA tissue IHC). |
| Staining is confined to cell membranes, extracellular material, or an unrelated structure. | That distribution conflicts with the reported nuclear and cytoplasmic pattern (HPA tissue IHC) and lack of a transmembrane segment (UniProt O00505: topology). Check morphology, counterstain, and detection controls before interpreting it as KPNA3 (standard IHC practice). |
| Strong staining appears in cells reported as Not detected. | HPA reports adipocytes, ovarian stroma cells, and splenic red-pulp cells as Not detected (HPA tissue IHC). Recheck cell identity and compare a no-primary control; cross-reactivity or endogenous detection activity can mimic a positive result (standard IHC practice). |
| Broad, hazy stain obscures nuclei and cell boundaries. | A diffuse haze cannot establish the reported cellular distribution (HPA tissue IHC: nuclear and cytoplasmic expression). Uneven reagent coverage, inadequate washing, or detection background are possible general IHC causes; inspect the no-primary control and the tissue edges (standard IHC practice). |
| No staining appears in an expected positive cell population. | A blank result in heart cardiomyocytes or adrenal glandular cells needs a technical check before biological interpretation (HPA tissue IHC: High in both). Check tissue preservation, retrieval conditions, primary-antibody application, and detection-system performance (standard IHC practice); no KPNA3-specific fixation sensitivity is supplied. |
💡Expected KPNA3 appearanceCall a positive result when identifiable nuclei, with possible cytoplasm, stain in an HPA High cell population such as heart cardiomyocytes; a membrane-only deposit or strong stain in HPA Not detected adipocytes is suspect (HPA tissue IHC; UniProt O00505: topology).
| Cell population and tissue | Use the named cells when judging intensity: HPA reports High staining in heart cardiomyocytes, colon endothelial cells, and duodenal glandular cells, but Not detected in adipocytes (HPA tissue IHC). A tissue-wide score can obscure these differences (standard IHC interpretation). |
| Strength of the tissue evidence | The tissue IHC assessment is Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC). Treat its cell-level observations as reference patterns, while resolving an unexpected result with controls and morphology (standard IHC practice). |
| Intracellular location and topology | UniProt lists cytoplasm and nucleus and reports no transmembrane segment (UniProt O00505: subcellular location and topology). These observations support intracellular staining and give no basis for calling an isolated membrane outline a KPNA3 pattern. |
| Antibody validation | HPA lists HPA046852 and HPA077643 as IHC Approved; HPA046852 is also ICC Supported (HPA antibodies). These statuses identify the reported application evidence; they do not establish that an untested antibody or every specimen will reproduce the pattern. |
| IF/ICC: where should KPNA3 appear? | Mainly in the nucleoplasm (HPA subcellular ICC-IF: supported), with possible cytosol (HPA subcellular ICC-IF: uncertain). This is a localization answer for IF/ICC, not an IHC-P staining-intensity standard. |
| Epitope and processing limits | UniProt reports no signal peptide or propeptide and lists a 2–521 chain (UniProt O00505: processing). The supplied sources give no antibody epitope or KPNA3-specific retrieval response, so neither a retrieval setting nor a fixation effect can be predicted from this record. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-control section is blank. | The assay may have failed at retrieval, primary-antibody application, or detection (standard IHC practice); HPA reports High staining in heart cardiomyocytes (HPA tissue IHC). | Confirm the section contains intact cardiomyocytes, then check reagent application and detection controls; optimize retrieval using the chosen antibody's IHC-P instructions (standard IHC practice). |
| Only a membrane rim is visible. | An isolated membrane pattern disagrees with the reported intracellular localization (HPA tissue IHC; UniProt O00505: topology). Deposited chromogen or nonspecific staining is possible (standard IHC practice). | Compare the no-primary control and inspect the rim at higher magnification. Score KPNA3 only where cellular staining matches the expected compartments (standard IHC practice; HPA tissue IHC). |
| Adipocytes stain strongly. | Adipocytes are reported as Not detected (HPA tissue IHC). Cell misidentification, nonspecific binding, or endogenous detection activity could explain the result (standard IHC practice). | Verify the cells on the counterstained section and compare no-primary and detection controls before assigning a KPNA3-positive score (standard IHC practice). |
| The whole section has diffuse brown haze. | Background may reflect excess detection signal, incomplete washing, or endogenous enzyme activity (standard IHC practice). Haze prevents assessment of the reported nuclear and cytoplasmic pattern (HPA tissue IHC). | Check the no-primary control, washing, and enzyme-blocking step; adjust the general detection workflow if the control also stains (standard IHC practice). |
| The nucleus and cytoplasm differ from the IF/ICC image. | HPA tissue IHC reports nuclear and cytoplasmic staining, while ICC-IF supports nucleoplasm and rates additional cytosol as uncertain (HPA tissue IHC; HPA subcellular ICC-IF). | Interpret the paraffin section against its IHC cell pattern and controls; use the ICC-IF result only as supporting localization evidence (HPA tissue IHC; HPA subcellular ICC-IF). |
| Staining varies across the section. | Uneven section coverage or processing can create a gradient (standard IHC practice). Cell populations also differ in reported KPNA3 staining (HPA tissue IHC). | Check whether the variation follows tissue anatomy or a reagent edge; compare the same identified cell type across well-preserved areas (standard IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing KPNA3 staining
A06131-1 · IHCImmunohistochemistry of paraffin-embedded rat stomach using KPNA3 Rabbit pAb at dilution of 1:100 .Perform high pressure antigen retrieval w
A06131 · IFImmunocytochemistry of KPNA3 in HeLa cells with KPNA3 antibody at 10 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000102753-KPNA3/tissue/cerebral+cortex#imid_20867111
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000102753-KPNA3/tissue/colon#imid_20867021
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000102753-KPNA3/tissue/liver#imid_20867038
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000102753-KPNA3/tissue/kidney#imid_20867031
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA046852; green = KPNA3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA046852; green = KPNA3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KPNA3 IHC & IF
🧪Run heart muscle first and look for staining in cardiomyocytes (HPA: High in cardiomyocytes). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); the supplied rows identify no verified negative cell population within heart muscle, so record cells without signal as unstained rather than assuming they are KPNA3-negative (HPA: heart muscle row).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KPNA3 in A-431, CACO-2, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), matched rabbit polyclonal IgG isotype, and KPNA3 knockout or immunizing-peptide block controls (datasheet: rabbit pAb; standard IHC practice). Quench endogenous peroxidase and check background in heart sections before interpreting chromogenic staining (standard IHC practice).
⚠️Feasibility: A KPNA3-specific fixation window or fixation effect is unreported, and the fixative in the A06131-1 paraffin-section caption is unreported (datasheet: A06131-1 caption). That caption uses high-pressure retrieval with 10 mM citrate at pH 6.0 and antibody at 1:100; it does not establish that retrieval is required (datasheet: A06131-1 caption). The supplied evidence does not establish whether frozen sections or IF are easier; assess endogenous peroxidase in heart muscle for chromogenic IHC and autofluorescence if using IF (standard IHC/IF practice).
HPA tissue IHC evidence for KPNA3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Section 3
Advanced KPNA3 IHC Tips
Troubleshoot KPNA3 staining in paraffin sections by checking retrieval, nuclear and cytoplasmic signal, cell identity, and controls before comparing scores.
Which retrieval conditions should I start with for KPNA3 in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate, pH 6.0, the condition reported for the catalog antibody in paraffin-embedded rat stomach (datasheet A06131-1). Record heating and cooling times, then compare adjacent sections processed together so a weak result can be assessed against the reported 1:100 staining condition (datasheet A06131-1; standard IHC practice). If staining remains weak, vary heating duration on matched sections before testing another retrieval buffer as a documented fallback (standard IHC practice). Score nuclear and cytoplasmic signal separately, since both patterns are reported in tissue (HPA: nuclear and cytoplasmic expression in most tissues).
Could fixation explain weak or uneven KPNA3 staining?
KPNA3-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (datasheet A06131-1). Record the specimen’s actual fixation conditions and compare matched sections under the same citrate pH 6.0 retrieval and antibody dilution before changing multiple steps (datasheet A06131-1; standard IHC practice). Examine whether weak staining tracks section thickness, folds, or uneven processing, and include a section with established positive staining in the same run (standard IHC practice). Do not assign a fixation effect from KPNA3 topology, modifications, or tissue expression; those records do not test fixation sensitivity (UniProt O00505; HPA: tissue IHC).
Should KPNA3 stain nuclei, cytoplasm, or both?
Assess nuclear and cytoplasmic staining separately: KPNA3 is annotated in both compartments, and tissue IHC reports both in most tissues (UniProt O00505: subcellular location; HPA: tissue IHC profile). Nucleoplasmic localisation has stronger subcellular support than cytosolic localisation in the supplied cell imaging record (HPA: nucleoplasm supported; cytosol uncertain). Use a nuclear counterstain and inspect individual cells at high magnification before calling diffuse brown signal cytoplasmic (standard IHC practice). If a section shows only extracellular deposits or signal restricted to damaged edges, check the negative control and tissue morphology before interpreting it as altered KPNA3 transport (standard IHC practice; UniProt O00505: nuclear import function).
How should I assess an unexpected KPNA3 epitope-dependent staining pattern?
The supplied record lists 0 isoforms, so it provides no basis for assigning two staining patterns to different KPNA3 isoforms (UniProt O00505: isoforms). KPNA3 has an IBB region at residues 2–58 and reported modifications at residues 2, 56, 60, and 484, but the supplied antibody evidence does not map its epitope (UniProt O00505: domains and modified residues; datasheet A06131-1). Compare matched sections after the stated citrate pH 6.0 retrieval and review antibody specificity documentation before attributing a lost signal to epitope masking (datasheet A06131-1; standard IHC practice). Keep nuclear and cytoplasmic observations distinct during that comparison (HPA: tissue IHC profile).
How can IF help investigate a disputed KPNA3 IHC pattern?
Use IF as a separate localisation check, since the supplied catalog evidence describes paraffin-section IHC rather than an IF protocol (datasheet A06131-1). Multiplex KPNA3 with a marker identifying the expected cell population, such as cardiomyocytes in heart muscle, and choose spectrally separated fluorophores after inspecting tissue autofluorescence (HPA: high in cardiomyocytes; standard IF practice). KPNA3 is intracellular with no transmembrane segment, so optimise permeabilisation for access to nuclear and cytoplasmic epitopes rather than assuming surface staining will suffice (UniProt O00505: topology and subcellular location; standard IF practice). Include single-stain and secondary-only controls when comparing channels and interpreting overlap (standard IF practice).
What should I check when KPNA3 DAB staining appears diffuse?
First compare a no-primary control with the stained section to identify signal caused by detection reagents or tissue rather than primary-antibody binding (standard IHC practice). For chromogenic detection, check the peroxidase block and limit DAB development while keeping both steps identical across comparison sections (standard IHC practice). Review blocking, washes, antibody concentration, and section drying; the reported 1:100 dilution is a starting condition for the selected rat stomach image, not a universal optimum (datasheet A06131-1; standard IHC practice). Preserve plausible nuclear and cytoplasmic staining when adjusting background, because both compartments are reported for KPNA3 (HPA: tissue IHC profile).
How should I score KPNA3 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then report nuclear and cytoplasmic results separately because KPNA3 staining occurs in both compartments (HPA: tissue IHC profile; standard IHC practice). For an H-score, multiply the percentage of cells in each intensity category by its category score; alternatively report % positive cells using one prespecified threshold (standard IHC practice). Normalise cell counts to the number of eligible cells, or positive-cell density to evaluated tissue area in mm², excluding folds and necrotic regions consistently (standard IHC practice). Keep retrieval, imaging, and scoring criteria matched across groups, and document the negative-control threshold (standard IHC practice).
When is an apparent KPNA3-positive cell or region convincing?
A convincing pattern follows intact cell boundaries and permits nuclear or cytoplasmic localisation, consistent with the supplied tissue profile and protein annotation (HPA: tissue IHC profile; UniProt O00505: subcellular location). Use cell identity as a check: high staining is reported in adrenal glandular cells and heart cardiomyocytes, whereas adipocytes are reported as not detected (HPA: tissue IHC cell-level observations). Treat staining confined to section edges, folds, necrotic areas, or the no-primary control as suspect, and check the peroxidase block for endogenous-enzyme signal (standard IHC practice). Interpret a disputed region alongside matched positive and negative controls before assigning a biological difference (standard IHC practice).
Boster reagents
Best KPNA3 / Importin subunit alpha-4 IHC Antibodies
The catalog provides KPNA3 images from paraffin-embedded rat stomach by IHC (A06131-1 image caption) and HeLa cells by ICC (A06131 image caption); both antibodies list human, mouse and rat reactivity (catalog reactivity).
Real IHC data
Anti-Importin subunit alpha-4 KPNA3 Antibody
Cat # A06131-1
Real IF data
Anti-Importin subunit alpha-4 KPNA3 Antibody
Cat # A06131
A06131-1 lists IHC and IF applications (catalog applications), with IHC shown in paraffin-embedded rat stomach at 1:100 after citrate retrieval (A06131-1 image caption). A06131 lists IF and ICC applications (catalog applications), with ICC shown in HeLa cells at 10 μg/mL (A06131 image caption).
Which to pick: Choose A06131-1 for paraffin-section IHC because its own image shows that preparation; the fixative is unreported (A06131-1 image caption). Choose A06131 for IF/ICC because those applications are listed and its own image shows ICC staining, while IHC is not listed (A06131 catalog applications; A06131 image caption). Both are rabbit antibodies with no clone specified (catalog host/clone), and both list human, mouse and rat reactivity (catalog reactivity); the pictured IHC evidence is specifically from rat tissue (A06131-1 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00505 (IMA4_HUMAN, Importin subunit alpha-4).
- Human Protein Atlas. KPNA3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KPNA3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. KPNA3 antibody validation summary (2 antibodies).
- SIRT1 promotes the progression and chemoresistance of colorectal cancer through the p53/miR-101/KPNA3 axis. Cancer biology & therapy 2023 — PMC10431729.
- KPNA3 Confers Sorafenib Resistance to Advanced Hepatocellular Carcinoma via TWIST Regulated Epithelial-Mesenchymal Transition. Journal of Cancer 2019 — PMC6692625.
- Exosomal Long Non-coding RNA HOTTIP Increases Resistance of Colorectal Cancer Cells to Mitomycin via Impairing MiR-214-Mediated Degradation of KPNA3. Frontiers in cell and developmental biology 2020 — PMC7876302.
- SIRT1 activation synergizes with FXR agonism in hepatoprotection via governing nucleocytoplasmic shuttling and degradation of FXR. Acta pharmaceutica Sinica. B 2023 — PMC9978964.
- PubMed PMID:9154134 — UniProt-cited evidence.
- PubMed PMID:9395085 — UniProt-cited evidence.
- PubMed PMID:9435235 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KPNA3 (Importin subunit alpha-4). Expected localisation Nuclear and cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A06131-1). Validated IHC antibody: A06131-1 at 1:50-1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat stomach tissue; fixative not specified (datasheet A06131-1); 2. Fixation: Image fixative and duration unreported (datasheet A06131-1); 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: Citrate pH 6.0 (datasheet A06131-1); 20 min, 95–100 °C (standard); 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-KPNA3, 1:50-1:100 (datasheet A06131-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06131-1); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No isoforms listed; mature chain spans residues 2–521 (UniProt); Regulation: Tissue enhanced in skeletal muscle and tongue (HPA tissue RNA). Evidence: UniProt O00505, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06131-1, PMC10431729, PMC6692625. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KPNA3/immunohistochemistry-ihc-guide.