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KPNB1 / Importin subunit beta-1 · IHC design guide
Design Immunohistochemistry for KPNB1
Plan KPNB1 chromogenic IHC in paraffin sections using the catalog antibody’s IHC-P conditions (datasheet: M01851-2). Compare cytoplasmic and nuclear staining with the broad tissue pattern and use glomerular cells as a high-staining reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KPNB1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M01851-2) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Liver+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). Selected-image fixative and duration unreported (datasheet M01851-2); verify before use. | |
| Caveat | Cholangiocytes may lack staining despite the broad pattern (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | Two isoforms; epitope differences are unspecified (UniProt) |
Section 1
Recommended KPNB1 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet M01851-2). One published KPNB1 IHC protocol is included (PMC9808660).
| Sample | Paraffin-embedded human thyroiditis tissue; fixative not specified (datasheet M01851-2) |
| Fixation | Image fixative and duration unreported (datasheet M01851-2); 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 M01851-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M01851-2) |
| Primary antibody | Mouse monoclonal (clone 3I11F2) anti-KPNB1, 2 μg/ml (datasheet M01851-2) |
| Primary incubation | Overnight at 4 °C (datasheet M01851-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M01851-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KPNB1-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet M01851-2); consider pH 9.0 when adapting the published protocol (PMC9808660).
Section 2
What Is the Expected KPNB1 Staining Pattern?
KPNB1 should stain the cytoplasm and nuclear region across many cell types in paraffin tissue sections (HPA: ubiquitous cytoplasmic and nuclear expression). Strong examples include glomerular cells, neuronal cells in the caudate, and pachytene spermatocytes (HPA: High). Interpret the pattern with some caution: tissue IHC reliability is Supported, with medium consistency against RNA data (HPA: reliability). KPNB1 has no transmembrane segment (UniProt Q14974: topology).
| Cytoplasmic and nuclear-region staining in glomerular cells or pachytene spermatocytes, with clear cellular boundaries. | This fits the reported distribution and High staining in those cells (HPA: tissue IHC). Compare intensity within the same run; High is an HPA category, not a universal chromogen threshold (HPA: tissue IHC; general IHC practice). |
| Signal confined to an extracellular edge or the cell surface, with no convincing cytoplasmic or nuclear-region component. | Treat the compartment mismatch as a possible artefact and review morphology and controls (HPA: cytoplasmic and nuclear tissue profile; UniProt Q14974: no transmembrane segment). A nuclear-rim emphasis is reported by ICC-IF, but need not be resolved in chromogenic tissue IHC (HPA: subcellular ICC-IF; general IHC practice). |
| Strong staining in liver cholangiocytes, parathyroid glandular cells, or vaginal squamous epithelial cells. | These specific cell populations are reported as Not detected (HPA: tissue IHC). Check cross-reactivity and, if using enzyme detection, endogenous enzyme activity before interpreting the signal as KPNB1 (general IHC practice). Do not treat the entire organ as negative (HPA: cell-specific tissue IHC). |
| Uniform haze extends through stroma, empty spaces, and cells with little morphological distinction. | This is less persuasive than a cell-associated cytoplasmic and nuclear-region pattern (HPA: tissue IHC; general IHC practice). Assess nonspecific background with a no-primary control and review blocking, antibody concentration, and wash conditions (general IHC practice). |
| No signal in glomerular cells or pachytene spermatocytes on an otherwise readable section. | Both are reported High, so an absent result calls for a technical review before a biological conclusion (HPA: tissue IHC; general IHC practice). Check section integrity, retrieval, primary and detection reagents, and the run's positive control (general IHC practice). |
💡Expected KPNB1 appearanceA convincing positive is cell-associated cytoplasmic and nuclear-region staining, readily visible in reported High glomerular cells or pachytene spermatocytes; staining confined to extracellular space or to HPA Not detected cell populations warrants control review (HPA: tissue IHC; general IHC practice).
| Tissue and cell selection (HPA: tissue IHC) | Use a reported High population, such as glomerular cells, to judge assay performance (HPA: High). Salivary glandular cells are Low, so faint staining there alone is a poor basis for declaring the run unsuccessful (HPA: Low; general IHC practice). |
| Antibody validation (HPA: antibody records) | HPA029878 and CAB034449 have Supported IHC status; HPA050302 has no IHC status in this record (HPA: antibodies). Match any validation claim to the antibody actually used, and interpret tissue staining alongside the profile's medium RNA–protein consistency (HPA: reliability). |
| Subcellular resolution (HPA: tissue IHC; HPA: ICC-IF) | Tissue IHC is described as cytoplasmic and nuclear (HPA: tissue IHC). ICC-IF reports an enhanced nuclear-membrane location with additional nucleoplasm and cytosol; that finer localisation does not set a required rim pattern for chromogenic sections (HPA: ICC-IF; general IHC practice). |
| Protein form and epitope (UniProt Q14974) | UniProt lists two isoforms, a single 1–876 chain, and no signal peptide or propeptide (UniProt Q14974: isoforms, processing). The supplied record gives no antibody epitope, so it cannot establish whether either isoform or a modified residue changes staining (UniProt Q14974: isoforms, modified residues; HPA: antibody records). |
| Antigen retrieval and detection controls (general IHC practice) | Retrieval conditions and enzyme-based detection can affect an IHC run generally (general IHC practice). No supplied KPNB1 source identifies a preferred retrieval condition or target-specific fixation sensitivity; evaluate changes with a positive tissue and appropriate controls (HPA: tissue IHC; general IHC practice). |
| IF/ICC Q: Where should signal appear? (HPA: subcellular ICC-IF) | A: Mainly at the nuclear membrane, with additional nucleoplasmic and cytosolic localisation (HPA: subcellular ICC-IF). This is the IF/ICC localisation reference; the primary application here is chromogenic IHC on paraffin sections (HPA: tissue IHC; HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Glomerular cells and pachytene spermatocytes both appear negative (HPA: High in each). | A shared assay failure is plausible when reported High populations lack signal (HPA: tissue IHC; general IHC practice). | Inspect morphology and the positive control; verify retrieval, primary-antibody incubation, and detection steps against the run record (general IHC practice). |
| Only a broad brown haze is visible, without cell-associated localisation (general IHC practice). | Nonspecific antibody binding, concentrated reagent, or insufficient washing may contribute (general IHC practice). | Review the no-primary control, blocking, antibody concentration, and washes; reassess whether signal follows the HPA cytoplasmic and nuclear pattern (general IHC practice; HPA: tissue IHC). |
| Cholangiocytes stain strongly despite their Not detected reference level (HPA: tissue IHC). | Cross-reactivity or endogenous enzyme activity may mimic a positive chromogenic result (general IHC practice). | Compare a no-primary control and an appropriate endogenous-enzyme control; identify cholangiocytes by morphology before scoring (general IHC practice; HPA: tissue IHC). |
| A Low-reference population is faint while a High-reference population stains clearly (HPA: tissue IHC). | The difference may reflect the reported cell-specific intensity rather than assay failure (HPA: High and Low categories). | Score each identified cell population against its own HPA category; retain the High-reference tissue as the run check (HPA: tissue IHC; general IHC practice). |
| Signal seems restricted to the cell edge or extracellular material (general IHC practice). | This conflicts with the reported cytoplasmic and nuclear-region distribution; edge staining can reflect artefact (HPA: tissue IHC; general IHC practice). | Recheck section morphology and controls, then compare staining with cytoplasm and the nuclear region in reported High cells (HPA: tissue IHC; general IHC practice). |
| A tissue section lacks a sharply defined nuclear rim, although cells show cytoplasmic and nuclear-region staining (HPA: tissue IHC). | A nuclear-membrane emphasis is an ICC-IF observation; chromogenic tissue IHC has different spatial resolution (HPA: subcellular ICC-IF; general IHC practice). | Judge the tissue section against HPA's cytoplasmic and nuclear IHC profile and its cell-specific levels; use the ICC-IF localisation only for the separate IF/ICC interpretation (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing KPNB1 staining
M01851-2 · IHCIHC analysis of KPNB1 using anti-KPNB1 antibody (M01851-2). KPNB1 was detected in a paraffin-embedded section of human thyroiditis tissue. H
M01851-2 · IHCIHC analysis of KPNB1 using anti-KPNB1 antibody (M01851-2). KPNB1 was detected in a paraffin-embedded section of human squamous cell lung ca
M01851-2 · IFIF analysis of KPNB1 using anti-KPNB1 antibody (M01851-2). KPNB1 was detected in an immunocytochemical section of CACO-2 cells. Enzyme antig
A01851-2 · IFIF analysis of KPNB1 using anti-KPNB1 antibody (A01851-2). KPNB1 was detected in an immunocytochemical section of CACO-2 cells. Enzyme antig
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108424-KPNB1/tissue/cerebral+cortex#imid_8371900
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108424-KPNB1/tissue/colon#imid_8371810
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108424-KPNB1/tissue/liver#imid_8371811
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108424-KPNB1/tissue/kidney#imid_8371812
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029878; green = KPNB1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029878; green = KPNB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KPNB1 IHC & IF
🧪Run kidney first: cells in glomeruli should stain (HPA: High in cells in glomeruli). Use liver cholangiocytes as the tissue negative (HPA: Not detected in cholangiocytes); cells without signal on the kidney slide should show counterstain alone, but the supplied HPA rows do not establish any neighboring cell type as KPNB1-negative.
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Liver (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KPNB1 in A-431, U-251MG, U2OS, CACO-2, HEK293, HeLa BAC 5737, HeLa , NIH 3T3, with annotated localisation: Nuclear membrane (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched mouse IgG isotype control (selected M01851-2 IHC caption: mouse primary antibody). Use KPNB1 knockout material as a biological negative, and block endogenous peroxidase when using HRP/DAB, particularly where blood remains in glomeruli (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M01851-2 tissue-IHC caption does not state the fixative. That paraffin-section caption used heat retrieval in EDTA at pH 8.0, so follow its retrieval conditions as a starting point; the supplied evidence does not establish whether frozen sections or IF/ICC are easier (selected M01851-2 IHC caption). Glomerular blood can contribute peroxidase background in chromogenic IHC, so assess the no-primary control alongside the kidney section (standard IHC practice).
HPA tissue IHC evidence for KPNB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Molecular layer cells - nucleus | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Testis | Pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced KPNB1 IHC Tips
Use compartment-resolved staining and matched controls to troubleshoot KPNB1 chromogenic IHC in paraffin sections; treat IF/ICC as a separate application (UniProt Q14974; HPA tissue IHC).
Which retrieval conditions should I start with for KPNB1 paraffin-section IHC?
Start with heat-mediated EDTA retrieval at pH 8.0 (datasheet M01851-2). The selected paraffin-section image used that retrieval before 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet M01851-2). Keep section thickness, heating time, cooling time, and detection conditions matched when comparing retrieval runs (standard IHC practice). If staining remains weak, test another retrieval buffer or heating duration on adjacent sections, with a known-positive control in every run (standard IHC practice). Score cytoplasmic and nuclear-rim signal separately, because KPNB1 has both cytoplasmic and nuclear-envelope annotations (UniProt Q14974 subcellular location).
How can I troubleshoot fixation when KPNB1 staining is weak or uneven?
The selected image identifies a paraffin section but gives no fixative or fixation duration; target-specific fixation sensitivity is therefore unknown (datasheet M01851-2). Record the fixative, time before fixation, specimen size, and fixation duration for each block before comparing staining intensity (standard IHC practice). Compare similarly processed sections with the same EDTA pH 8.0 retrieval and detection run to limit processing differences (datasheet M01851-2; standard IHC practice). Patchy loss alongside poor tissue preservation warrants a repeat on a better-preserved block (standard IHC practice). Do not assign a KPNB1-specific fixation effect from its distribution or modifications alone (HPA tissue IHC; UniProt Q14974 modified residues).
What KPNB1 staining pattern should count as plausible in paraffin sections?
Evaluate cytoplasmic signal and signal at the nuclear envelope as plausible KPNB1 compartments (UniProt Q14974 subcellular location). HPA describes mainly nuclear-membrane localisation, with additional nucleoplasmic and cytosolic signal, and reports ubiquitous cytoplasmic and nuclear tissue staining (HPA subcellular; HPA tissue IHC). Use a nuclear counterstain and sufficient magnification to distinguish a nuclear rim from diffuse nuclear chromogen (standard IHC practice). Compare cells within the same compartment and tissue region, because cell-level expression differs across tissues (HPA tissue IHC). Diffuse staining over acellular material or a section-wide rim should prompt control review before being scored as KPNB1 (standard IHC practice).
Could isoforms or epitope accessibility explain discordant KPNB1 IHC results?
KPNB1 has 2 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform-specific staining (UniProt Q14974 isoforms; datasheet M01851-2). Request the immunogen region before interpreting discordant antibodies as isoform differences (standard IHC practice). The protein has an importin N-terminal domain at residues 21–101 and annotated modified residues at 12, 211, 835, and 867; their effect on this antibody is untested here (UniProt Q14974 domains and modified residues; datasheet M01851-2). Compare adjacent sections under identical retrieval and detection conditions (standard IHC practice). Treat altered staining as an epitope-accessibility hypothesis until independently validated (standard IHC practice).
How should I adapt KPNB1 localisation checks to multiplex IF/ICC?
Use IF/ICC as a separate validation experiment, since the selected antibody evidence here documents paraffin-section chromogenic IHC (datasheet M01851-2). Multiplex KPNB1 with a marker identifying the expected cell population, and include single-stain controls to check channel bleed-through (standard IF practice). Choose fluorophores in channels with low tissue autofluorescence and assess an unstained specimen before interpreting faint signal (standard IF practice). KPNB1 has no transmembrane segment and is annotated in the cytoplasm and nuclear-envelope region, so permeabilisation should allow antibody access to intracellular epitopes (UniProt Q14974 topology and subcellular location; standard IF practice). Optimise permeabilisation and antibody conditions for IF/ICC independently (standard IF practice).
What controls help separate KPNB1 signal from chromogenic background?
Include a no-primary control and inspect its DAB deposition before assigning weak brown staining to KPNB1 (standard IHC practice). Block endogenous peroxidase for an HRP/DAB workflow, particularly when background follows tissue structures rather than cell boundaries (standard IHC practice). The selected image used 10% goat serum, 2 μg/ml mouse primary antibody overnight at 4°C, and a peroxidase-conjugated anti-mouse secondary for 30 minutes at 37°C (datasheet M01851-2). If background persists, compare a primary-antibody titration and shorter chromogen development using matched sections (standard IHC practice). Preserve enough counterstain to judge cytoplasmic and nuclear-rim distribution (UniProt Q14974 subcellular location; standard IHC practice).
How should KPNB1 IHC be scored across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions and cell populations before scoring, and exclude folds, necrosis, and section edges using the same rules for every specimen (standard IHC practice). For cellular staining, report the percentage of positive cells and an H-score from intensity categories; record cytoplasmic and nuclear-rim scores separately (standard IHC practice; UniProt Q14974 subcellular location). Normalise each score to the number of evaluable cells in its defined population, rather than total tissue area (standard IHC practice). If counting discrete positive cells, report density per mm² of evaluable tissue and the analysed area (standard IHC practice). Keep retrieval, DAB development, imaging, and thresholds consistent across comparisons (standard IHC practice).
When should apparent KPNB1 positivity be treated as artefact?
Interpret cell-associated cytoplasmic or nuclear-rim staining as plausible only when controls and tissue morphology support it (UniProt Q14974 subcellular location; standard IHC practice). HPA reports high staining in caudate neuronal cells and kidney glomerular cells, while liver cholangiocytes are listed as not detected; use such cell-level patterns as context, not an absolute diagnostic rule (HPA tissue IHC). Signal confined to an implausible compartment, necrotic area, fold, or section edge requires review of adjacent tissue and controls (standard IHC practice). Brown staining in a no-primary section raises concern for endogenous enzyme activity or detection background (standard IHC practice). Confirm disputed patterns with matched controls and independent validation (standard IHC practice).
Boster reagents
Best KPNB1 / Importin subunit beta-1 IHC Antibodies
Anti-KPNB1 antibodies have IHC data from human paraffin sections and IF data from CACO-2 cells (catalog image captions); both list human, mouse and rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-KPNB1 Antibody ® (monoclonal, 3I11F2)
Cat # M01851-2
Real IF data
Anti-KPNB1 Antibody ®
Cat # A01851-2
M01851-2 has IHC images from human thyroiditis, squamous cell lung carcinoma, renal carcinoma and breast cancer paraffin sections (M01851-2 image captions). A01851-2 has IF data from CACO-2 cells (A01851-2 image caption).
Which to pick: Choose M01851-2 for tissue IHC: its application list includes IHC, and its images show paraffin sections with EDTA pH 8.0 heat retrieval; the fixative is unreported (M01851-2 application list and IHC image captions). Choose A01851-2 for IF/ICC in CACO-2 cells; its application list excludes IHC (A01851-2 application list and IF image caption). Both list human, mouse and rat reactivity, but the supplied IHC and IF images do not demonstrate staining in mouse or rat samples (catalog reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14974 (IMB1_HUMAN, Importin subunit beta-1).
- Human Protein Atlas. KPNB1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. KPNB1 subcellular location (ICC-IF): Mainly localized to the nuclear membrane. In addition localized to the nucleoplasm and cytosol..
- Human Protein Atlas. KPNB1 antibody validation summary (3 antibodies).
- Expression of Karyopherin Alpha 2 and Karyopherin Beta 1 Correlate with Poor Prognosis in Gastric Cancer. Oncology 2022 — PMC9808660.
- EZH2-miR-30d-KPNB1 pathway regulates malignant peripheral nerve sheath tumour cell survival and tumourigenesis. The Journal of pathology 2014 — PMC4166508.
- Multi-omics analysis identifies a major histocompatibility complex class II-associated antigen-presenting cancer-associated fibroblast-like state linked to the nuclear factor erythroid 2-related factor 2-karyopherin subunit beta 1 axis in nonsmall cell lung cancer. Journal of cell communication and signaling 2026 — PMC13238553.
- KPNB1-ATF4 induces BNIP3-dependent mitophagy to drive odontoblastic differentiation in dental pulp stem cells. Cellular & molecular biology letters 2024 — PMC11600598.
- PubMed PMID:7615630 — UniProt-cited evidence.
- PubMed PMID:7627554 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KPNB1 (Importin subunit beta-1). Expected localisation Cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M01851-2). Validated IHC antibody: M01851-2 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Liver. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human thyroiditis tissue; fixative not specified (datasheet M01851-2); 2. Fixation: Image fixative and duration unreported (datasheet M01851-2); 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 M01851-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M01851-2); 8. Primary antibody: Mouse monoclonal (clone 3I11F2) anti-KPNB1, 2 μg/ml (datasheet M01851-2); 9. Primary incubation: Overnight at 4 °C (datasheet M01851-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet M01851-2); verify before use.; Caveat: Cholangiocytes may lack staining despite the broad pattern (HPA tissue IHC); Isoform / epitope: Two isoforms; epitope differences are unspecified (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt Q14974, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nuclear membrane. In addition localized to…, Datasheet M01851-2, PMC9808660. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KPNB1/immunohistochemistry-ihc-guide.