This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
KCNMB4 / Calcium-activated potassium channel subunit beta-4 · IHC design guide
Design Immunohistochemistry for KCNMB4
Plan KCNMB4 paraffin IHC around high Purkinje cell cytoplasm/membrane staining (HPA tissue IHC). The IHC-validated antibody has a 1:100–1:300 dilution range (datasheet); cerebellum and adipose tissue offer reported positive and negative staining examples, respectively (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KCNMB4 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 | Purkinje cell cytoplasm/membrane (HPA tissue IHC) | |
| Staining pattern | High Purkinje cytoplasm/membrane; medium glia and tubules (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Brain-predominant expression (UniProt) | |
| Isoform / epitope | 0 isoforms annotated; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended KCNMB4 IHC & IF Protocols
The catalog antibody protocol is accompanied by one published KCNMB4 IHC method using FFPE sections (PMC12789566).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A06117) |
| Fixation | Image fixative and duration unreported (datasheet A06117); 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-KCNMB4, 1:100-1:300 (datasheet A06117) |
| 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 | KCNMB4-positive staining in purkinje cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Expression in CNS, adrenal gland and kidney. 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 setting); the article does not report retrieval conditions (PMC12789566).
Section 2
What Is the Expected KCNMB4 Staining Pattern?
In paraffin-section IHC, expect the clearest KCNMB4 staining in cerebellar Purkinje cells, with cytoplasmic and membrane signal (HPA: High; IHC reliability Enhanced). Glial cells in cerebral cortex and hippocampus show medium staining (HPA: Medium). KCNMB4 is a membrane protein with two transmembrane segments and cytoplasmic termini (UniProt Q86W47 topology). HPA also reports cytosolic localisation by ICC-IF, so interpret compartment patterns in the context of the assay (HPA: Cytosol, approved).
| Strong Purkinje-cell cytoplasmic or membrane staining in cerebellum, with identifiable cell outlines. | This matches the strongest supplied tissue-IHC observation (HPA: Purkinje cells, High; cytoplasm/membrane). UniProt places KCNMB4 in the membrane (UniProt Q86W47 topology). Score the stained cells and compartment together; cytoplasmic signal within Purkinje cells is part of the reported IHC pattern and does not, by itself, indicate an artefact. |
| Moderate staining in cortical or hippocampal glial cells, or in kidney proximal-tubule cell bodies. | These are reported positive populations at medium intensity (HPA: cerebral-cortex glial cells, hippocampal glial cells, kidney proximal tubules). They can support an interpretation made in cerebellum, but do not require every cell in a section to stain. Compare the observed cell type with the tissue annotation before treating a regional signal as KCNMB4. |
| Predominantly nuclear staining, without the expected Purkinje-cell cytoplasmic or membrane pattern. | Treat a nuclear-only pattern as suspect: the supplied IHC annotation is cytoplasm/membrane and UniProt describes a membrane protein (HPA: Purkinje cells; UniProt Q86W47 topology). Review the unstained morphology, counterstain and detection controls under general IHC practice. The sources do not establish a target-specific reason for nuclear staining. |
| Strong signal in a cell population annotated as unstained, or staining that covers much of the section. | A mismatched cell pattern can reflect antibody cross-reactivity or endogenous detection activity (general IHC practice). For example, HPA reports adipocytes in adipose tissue and hematopoietic cells in bone marrow as not detected (HPA: tissue IHC). Those observations are useful comparators, not proof that every cell in those tissues is KCNMB4-free. |
| No discernible signal in Purkinje cells despite preserved tissue morphology. | The result fails to reproduce the supplied high-level positive reference (HPA: cerebellar Purkinje cells, High). First check the IHC run and its positive control using general IHC practice. A blank section alone cannot distinguish assay failure from specimen variation; HPA rates overall tissue-IHC reliability Enhanced but notes only medium consistency with RNA expression. |
💡Expected KCNMB4 appearanceCall a convincing positive result strong Purkinje-cell cytoplasmic or membrane staining (HPA: High), with medium staining possible in the other annotated cell populations (HPA: tissue IHC); isolated nuclear staining or broad signal in annotated negative cells is suspect under general IHC practice.
| Assay and compartment | Tissue IHC records Purkinje-cell cytoplasm/membrane staining, whereas ICC-IF records cytosol (HPA: tissue IHC; HPA: Cytosol, approved). Compare each observation within its assay; the ICC-IF entry does not establish how a paraffin section must look. |
| Protein topology | KCNMB4 has two transmembrane segments, an extracellular region at residues 41–167 and cytoplasmic termini (UniProt Q86W47 topology). The supplied evidence gives no antibody epitope, so topology cannot establish which part this stain detects or predict retrieval needs. |
| Tissue and antibody evidence | Brain-enhanced RNA expression and predominant brain expression support choosing cerebellum as a reference (HPA: tissue RNA; UniProt Q86W47 tissue specificity). The listed antibody has Enhanced IHC validation (HPA: HPA072287); HPA still describes medium consistency between staining and RNA data. |
| Protein glycosylation | Two glycosylation sites are annotated at residues 53 and 90 (UniProt Q86W47). Their effect on this antibody's staining is unreported in the supplied evidence; do not infer a retrieval method, intensity change or tissue-specific failure from those sites. |
| Situation | Likely cause | Next action |
|---|---|---|
| Purkinje cells are blank in an otherwise readable cerebellar section. | The expected high-level reference is missing (HPA: Purkinje cells, High); the evidence does not identify a KCNMB4-specific failure mechanism. | Check a known-positive section, primary-antibody setup, retrieval record and chromogenic detection run (general IHC practice). Compare morphology and control performance before calling the sample negative. |
| Only nuclei stain in cerebellum. | This compartment conflicts with the reported Purkinje-cell cytoplasm/membrane pattern (HPA: tissue IHC) and membrane topology (UniProt Q86W47). | Recheck counterstain interpretation and detection controls, then repeat with an appropriate positive control if needed (general IHC practice). Do not score nuclear-only signal as the expected pattern. |
| Brown colour is diffuse across cells and extracellular areas. | Poorly localised colour can arise from background or endogenous detection activity (general IHC practice); it does not reproduce the cell-resolved HPA pattern. | Inspect a no-primary control and review blocking, washing and detection conditions (general IHC practice). Interpret only signal that resolves to annotated cells and compartments (HPA: tissue IHC). |
| Adipocytes or marrow hematopoietic cells stain strongly. | Those populations are annotated as not detected (HPA: adipose tissue; bone marrow). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Compare a no-primary control and a cerebellar positive section in the same run (general IHC practice; HPA: Purkinje cells, High). Treat a negative-tissue mismatch as a specificity question, not definitive proof of absence. |
| Cortex or hippocampus stains, but mainly outside glial cells. | The reported medium-level populations are glial cells in both regions (HPA: cerebral cortex; hippocampus); another dominant cell pattern needs separate assessment. | Confirm cell identity from morphology and compare the cerebellar pattern and controls (general IHC practice; HPA: Purkinje cells, High). Avoid scoring all regional colour as a validated KCNMB4-positive cell type. |
| An ICC-IF image looks cytosolic while the paraffin IHC section shows membrane-associated Purkinje staining. | HPA reports approved cytosolic localisation in ICC-IF and cytoplasm/membrane staining in tissue IHC (HPA: subcellular; tissue IHC). | Evaluate each image against its own assay annotation (HPA: ICC-IF; tissue IHC). Use the separate IF/ICC guide for that assay; this IHC pattern alone does not resolve the compartment difference. |
Experiment evidence
IHC & IF images showing KCNMB4 staining
A06117 · IHCImmunohistochemistry analysis of MaxiKβ antibody in paraffin-embedded human brain tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000135643-KCNMB4/tissue/cerebral+cortex#imid_20025457
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000135643-KCNMB4/tissue/colon#imid_20025959
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000135643-KCNMB4/tissue/liver#imid_20025976
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000135643-KCNMB4/tissue/kidney#imid_20025969
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA072287; green = KCNMB4, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA072287; green = KCNMB4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KCNMB4 IHC & IF
🧪Run cerebellum first and require staining in Purkinje cells (High; HPA: cerebellum, Purkinje cells, cytoplasm/membrane). Use adipose tissue as the negative tissue (HPA: adipocytes, Not detected); on the cerebellum slide, neighboring non-Purkinje cells should lack staining comparable to the Purkinje-cell signal.
Positive control tissue: Cerebellum (Purkinje cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KCNMB4 in RT-4, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, an isotype control matched to the primary antibody’s host species and clonality, and KCNMB4-knockout tissue if available (standard IHC controls). Block endogenous peroxidase for chromogenic detection and check cerebellar neuronal pigment against the no-primary slide (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected A06117 paraffin-section brain caption also leaves the fixative unreported (selected tissue-IHC caption). Antigen-retrieval dependence must be determined empirically, and the supplied evidence does not establish whether frozen sections or IF are easier for KCNMB4 (standard IHC practice; selected tissue-IHC caption). Cerebellar neuronal pigment may complicate chromogenic scoring, so compare the signal with the no-primary control (standard IHC practice).
HPA tissue IHC evidence for KCNMB4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Medium | Protein (IHC) | HPA → |
| Kidney | Proximal tubules (cell body) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KCNMB4 IHC Tips
Troubleshoot KCNMB4 staining in paraffin sections by checking retrieval, compartment, cell type and controls before comparing chromogenic signal across samples.
What retrieval should I start with for weak KCNMB4 staining in paraffin sections?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval specification). KCNMB4 has an extracellular region at residues 41–167 between two transmembrane segments, so epitope accessibility may depend on antibody binding site (UniProt Q86W47 topology). If staining is weak, test a stronger retrieval condition on adjacent sections while holding antibody concentration and detection time constant (standard IHC practice). Compare the same cell population across conditions, using Purkinje cells as a reference when cerebellum is available (HPA: high cytoplasmic/membrane staining in Purkinje cells). Excessive retrieval can damage section morphology or increase background, so retain the mildest condition giving interpretable staining (standard IHC practice).
How should I troubleshoot possible fixation effects on KCNMB4 IHC?
The selected antibody image shows KCNMB4 staining in paraffin-embedded human brain, but its caption does not state the fixative (A06117 tissue-IHC caption). Target-specific fixation sensitivity is therefore unknown; record the actual fixative, fixation duration and processing history for each specimen before comparing results (A06117 tissue-IHC caption; standard IHC practice). On adjacent sections, keep retrieval at citrate pH 6.0, 95–98 °C for 20 min while changing only one downstream staining condition at a time (page retrieval specification; standard IHC practice). Check morphology and a process control before attributing weak signal to fixation, since poor preservation can also make cell-level scoring unreliable (standard IHC practice).
Should KCNMB4 staining be membranous or cytoplasmic in brain sections?
KCNMB4 is annotated as a membrane protein with transmembrane segments at residues 20–40 and 168–188 (UniProt Q86W47 topology). In cerebellar tissue IHC, Purkinje cells show high cytoplasmic/membrane staining, so assess both patterns within morphologically identified cells (HPA: Purkinje cells, high). HPA also reports an approved cytosolic location from subcellular imaging, which does not by itself establish the expected distribution in every paraffin section (HPA subcellular: cytosol). Record membrane-associated and diffuse cytoplasmic signal separately, and compare each with a no-primary control and adjacent intact tissue (standard IHC practice). Diffuse staining across unrelated structures deserves further validation before assignment to KCNMB4 (UniProt Q86W47: predominantly brain; standard IHC practice).
How can epitope position change the KCNMB4 IHC result?
The supplied record lists one 210-aa KCNMB4 chain and no annotated isoforms, so isoform-specific staining cannot be inferred from these data (UniProt Q86W47 record). Its N-terminal residues 1–19 and C-terminal residues 189–210 are cytoplasmic, while residues 41–167 are extracellular (UniProt Q86W47 topology). Two glycosylation sites, at residues 53 and 90, lie in that extracellular region and could affect access to an overlapping epitope; the antibody epitope is unspecified here (UniProt Q86W47 glycosylation and topology). Ask for the immunogen or mapped epitope before interpreting retrieval sensitivity, and compare staining with a second independently validated epitope when available (standard IHC validation practice).
How should I check a KCNMB4 IHC pattern by tissue immunofluorescence?
Use a marker that identifies the cell population being scored, such as a validated Purkinje-cell marker in cerebellum, alongside KCNMB4 (HPA: high Purkinje-cell staining; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue in each channel before assigning weak signal, because tissue autofluorescence can mimic antibody staining (standard IF practice). KCNMB4 has cytoplasmic termini and an extracellular loop; set permeabilisation according to the antibody’s mapped epitope, which is not supplied here (UniProt Q86W47 topology). Titrate detergent gently if an intracellular epitope requires access, and include single-label and no-primary controls to assess bleed-through and background (standard IF practice). Treat IF localisation as a cross-check of cell identity and compartment, then score chromogenic IHC on its own controls (standard IHC/IF practice).
What should I change if KCNMB4 chromogenic staining is diffuse?
First compare the section with a no-primary control and inspect whether color develops in the same structures, which points to detection or tissue background (standard IHC practice). Include a peroxidase block before peroxidase-based detection and optimise protein blocking, washes and primary-antibody concentration one variable at a time (standard chromogenic IHC practice). Avoid lengthening DAB development simply to recover faint signal, because extended development can obscure cell boundaries and raise nonspecific color (standard chromogenic IHC practice). Prioritise intact cerebellar Purkinje cells when available, where high cytoplasmic/membrane staining is reported, rather than interpreting uniform staining across all cells as expected KCNMB4 distribution (HPA: Purkinje cells, high; UniProt Q86W47: predominantly brain).
How should I quantify KCNMB4 staining across paraffin brain sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring; cerebellar Purkinje cells and cortical or hippocampal glial cells have different reported staining levels (HPA: Purkinje cells high; cortical and hippocampal glial cells medium). Within each region, report the percentage of positive cells plus an intensity-based H-score using categories 0–3, with a fixed positivity threshold across the batch (standard IHC scoring practice). Normalise positive-cell counts to all evaluable cells of the specified type, or report positive-cell density per mm² of viable tissue (standard IHC quantification practice). Exclude folds, torn edges and necrosis consistently, and document retrieval, detection and imaging settings before comparing sections (standard IHC practice).
How can I distinguish true KCNMB4 staining from artefact?
A plausible result has reproducible staining in an identified cell population, with cerebellar Purkinje-cell cytoplasmic/membrane staining providing a reference pattern (HPA: Purkinje cells, high). KCNMB4 is a membrane subunit, while HPA also reports cytosolic localisation, so compartment alone cannot prove identity; scrutinise nuclear-only or structure-free diffuse color (UniProt Q86W47: membrane; HPA subcellular: cytosol; standard IHC interpretation). Signal concentrated at cut edges, folds or necrotic areas should be excluded, and staining retained in a no-primary control warrants investigation of endogenous enzyme activity or detection background (standard chromogenic IHC practice). Confirm a candidate pattern across intact sections and an independent specificity control before making a biological claim, particularly outside the predominantly brain expression reported by UniProt (UniProt Q86W47 tissue specificity; standard IHC validation practice).
Boster reagents
Best KCNMB4 / Calcium-activated potassium channel subunit beta-4 IHC Antibodies
A06117 has an IHC image from paraffin-embedded human brain tissue (catalog image caption). IF is listed as an application without an IF image (catalog applications; image inventory); reactivity lists human, mouse and rat (catalog reactivity).
Real IHC data
Anti-MaxiK beta KCNMB4 Antibody
Cat # A06117
A06117 will render with its human brain paraffin-section IHC figure (catalog image caption). IF is listed without a corresponding figure, and human, mouse and rat are listed as reactive species (catalog applications; image inventory; reactivity).
Which to pick: For tissue IHC, choose A06117: its own image shows paraffin-embedded human brain tissue, and the listed IHC dilution is 1:100–1:300 (catalog image caption; datasheet: IHC dilution). The caption does not report the fixative (catalog image caption). For IF/ICC planning, A06117 lists IF at 1:50, but has no IF image or ICC application listed; for mouse or rat work, reactivity is listed without a species-specific IHC figure (datasheet: IF dilution; catalog applications, reactivity and image inventory).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q86W47 (KCMB4_HUMAN, Calcium-activated potassium channel subunit beta-4).
- Human Protein Atlas. KCNMB4 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. KCNMB4 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. KCNMB4 antibody validation summary (1 antibodies).
- PRMT5 upregulates KCNMB4 expression via histone methylation to promote paclitaxel resistance in advanced nasopharyngeal carcinoma. Cell death & disease 2026 — PMC12789566.
- Recurrent LRP1-SNRNP25 and KCNMB4-CCND3 fusion genes promote tumor cell motility in human osteosarcoma. Journal of hematology & oncology 2014 — PMC4197299.
- PubMed PMID:10792058 — UniProt-cited evidence.
- PubMed PMID:10828459 — UniProt-cited evidence.
- PubMed PMID:10692449 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KCNMB4 (Calcium-activated potassium channel subunit beta-4). Expected localisation Purkinje cell cytoplasm/membrane (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06117 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells - cytoplasm/membrane); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A06117); 2. Fixation: Image fixative and duration unreported (datasheet A06117); 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-KCNMB4, 1:100-1:300 (datasheet A06117); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: Brain-predominant expression (UniProt). Evidence: UniProt Q86W47, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet A06117, PMC12789566. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KCNMB4/immunohistochemistry-ihc-guide.