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KCNQ5 / Potassium voltage-gated channel subfamily KQT member 5 · IHC design guide
Design Immunohistochemistry for KCNQ5
Plan KCNQ5 paraffin IHC using skeletal muscle myocytes and cortical neurons as reference positives (HPA tissue IHC). Follow the catalog antibody’s documented IHC conditions (datasheet A04705-1), and interpret cytoplasmic staining (HPA tissue IHC) alongside the annotated membrane location (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KCNQ5 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); cell membrane expected (UniProt) | |
| Staining pattern | Cytoplasmic in myocytes and cortical neurons (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04705-1) | |
| Positive control | Skeletal muscle+1 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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Low agreement between staining and RNA (HPA tissue IHC) | |
| Regulation | No staining-linked regulator reported (UniProt) | |
| Isoform / epitope | 7 isoforms; check epitope isoform and membrane side (UniProt) |
Section 1
Recommended KCNQ5 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A04705-1) with a published paraffin-section protocol for guinea pig eyes (PMC8733613).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04705-1) |
| Fixation | Image fixative and duration unreported (datasheet A04705-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04705-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04705-1) |
| Primary antibody | Rabbit anti-KCNQ5, 2-5μg/ml (datasheet A04705-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04705-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04705-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KCNQ5-positive staining in myocytes of skeletal muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, most abundant in skeletal muscle. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A04705-1); the published protocol does not report retrieval (PMC8733613).
Section 2
What Is the Expected KCNQ5 Staining Pattern?
KCNQ5 is a six-pass cell-membrane channel with cytoplasmic regions (UniProt Q9NR82 topology). In paraffin-section IHC, expect the strongest observed staining in skeletal-muscle myocytes and moderate staining in cerebral-cortex neuronal cells; HPA describes the tissue pattern as cytoplasmic (HPA tissue IHC). Treat that appearance as a working expectation: HPA rates the IHC antibody Approved but reports low consistency between staining and RNA expression (HPA tissue IHC; HPA016655 IHC).
| Myocytes stain strongly, while cerebral-cortex neuronal cells stain more moderately (HPA tissue IHC). | This matches the reported positive-tissue contrast: skeletal muscle is High and cerebral cortex is Medium (HPA tissue IHC). Compare the intended cells within each section; staining throughout unrelated tissue compartments is less persuasive than this cell-specific pattern (general IHC practice). |
| Signal appears cytoplasmic, with possible membrane-associated emphasis in the expected cells. | HPA describes cytoplasmic tissue staining, while UniProt places the six-pass channel at the cell membrane and annotates cytoplasmic regions (HPA tissue IHC; UniProt Q9NR82 topology). Do not require a sharp membrane outline to call a paraffin-section result consistent with the reported IHC pattern. |
| Prominent staining appears in cells reported as unstained, such as adipocytes or cerebellar granular-layer cells (HPA tissue IHC). | That distribution conflicts with these HPA observations and may indicate cross-reactivity or endogenous detection activity; it does not establish either cause by itself (HPA tissue IHC; general IHC practice). Review a no-primary control and the staining distribution before assigning the signal to KCNQ5. |
| Diffuse chromogen covers many cells or the section without a clear cell-specific pattern. | Broad background makes localisation and intensity difficult to score (general IHC practice). Compare the no-primary control, inspect blocking and detection steps, and reassess primary-antibody concentration; these are general IHC checks, not evidence of a KCNQ5-specific fixation effect. |
| Expected myocytes show no detectable signal in skeletal muscle (HPA tissue IHC). | A blank known-positive tissue makes a negative result elsewhere hard to interpret (general IHC practice). Check tissue integrity, antibody application, retrieval conditions and detection controls before concluding that the experimental tissue lacks KCNQ5; HPA reports High myocyte staining but limited staining–RNA consistency (HPA tissue IHC). |
💡Expected KCNQ5 appearanceCall a result consistent when skeletal-muscle myocytes show strong, predominantly cytoplasmic staining and cerebral-cortex neuronal cells show more moderate staining; broad signal in reported negative cells or controls argues against a specific positive call (HPA tissue IHC; general IHC practice).
| Cellular compartment (UniProt Q9NR82 topology; HPA tissue IHC) | UniProt annotates cell-membrane localisation and six transmembrane segments; HPA reports cytoplasmic tissue staining (UniProt Q9NR82 topology; HPA tissue IHC). Interpret membrane-associated detail alongside the observed cytoplasmic pattern, rather than treating either description as a complete map of chromogenic signal. |
| Tissue and cell choice (HPA tissue IHC) | Skeletal-muscle myocytes are High, cerebral-cortex neuronal cells Medium, and hippocampal neuronal cells Low (HPA tissue IHC). Use the first two as positive-pattern references; weak hippocampal staining alone offers less leverage for judging whether an assay worked. |
| Antibody evidence (HPA016655 IHC; HPA tissue IHC) | The listed antibody is IHC Approved, while HPA reports low consistency between staining and RNA expression (HPA016655 IHC; HPA tissue IHC). Approval supports using the observed pattern as a reference, but a new tissue result still needs appropriate controls. |
| Isoform coverage (UniProt Q9NR82 isoforms) | UniProt lists seven KCNQ5 isoforms; this payload gives no antibody epitope or isoform coverage (UniProt Q9NR82 isoforms). A negative section cannot be assigned to a particular isoform from these records. Check epitope information if isoform coverage matters to the experiment. |
| IF/ICC Q&A: Should cytoplasmic bodies define the IHC result? | No. HPA calls its ICC-IF cytoplasmic-body location uncertain, and the listed antibody's ICC status is Uncertain (HPA subcellular ICC-IF; HPA016655 ICC). Use the separate IF/ICC guide for that application; the paraffin IHC tissue pattern is the reference here (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Skeletal-muscle myocytes are blank despite an expected High result (HPA tissue IHC). | The cause is unresolved from the slide alone; tissue preparation, retrieval, primary-antibody application or detection may need review (general IHC practice). | Run a documented positive section alongside the sample, verify each staining step and review retrieval conditions (general IHC practice). Do not infer KCNQ5-specific fixation sensitivity: none is supplied. |
| Cerebral cortex looks weaker than skeletal muscle. | A difference can match the reported Medium neuronal versus High myocyte staining (HPA tissue IHC). | Score the expected cells and compare matched runs before adjusting the assay (general IHC practice). Escalate only if the cortex is absent despite a working positive control; the HPA levels describe observations, not a required intensity ratio. |
| Reported negative cells stain, such as adipocytes or adrenal glandular cells (HPA tissue IHC). | Possible explanations include nonspecific binding or endogenous detection activity; the unexpected signal alone cannot distinguish them (general IHC practice). | Inspect no-primary and negative-tissue controls, then review blocking and detection reagents (general IHC practice). Record whether the signal follows the expected cells before scoring it as KCNQ5. |
| Diffuse chromogen obscures cell boundaries. | Excess background can arise during primary-antibody or detection steps (general IHC practice). | Compare the no-primary control, review blocking, and adjust antibody concentration or detection conditions according to the validated assay (general IHC practice). Reassess whether myocytes and neuronal cells remain distinguishable (HPA tissue IHC). |
| Nuclear-only signal dominates the expected positive cells. | Nuclear-only localisation conflicts with HPA's cytoplasmic tissue profile and UniProt's cell-membrane annotation (HPA tissue IHC; UniProt Q9NR82 subcellular location). | Treat the result as unresolved: inspect controls, tissue morphology and antibody conditions before calling it positive (general IHC practice). Seek independent support if nuclear localisation is central to the study. |
| A low-staining tissue is the only apparent negative result. | HPA reports Low hippocampal neuronal staining and Not detected cerebellar granular-layer staining; either can yield little visible signal (HPA tissue IHC). | Check skeletal-muscle myocytes in the same run before interpreting the negative sample (HPA tissue IHC; general IHC practice). Document cell type and control outcome rather than treating low signal as proof that the assay failed. |
Experiment evidence
IHC & IF images showing KCNQ5 staining
A04705-1 · IHCIHC analysis of KCNQ5 using anti-KCNQ5 antibody (A04705-1). KCNQ5 was detected in paraffin-embedded section of mouse brain tissue. Heat medi
A04705-1 · IHCIHC analysis of KCNQ5 using anti-KCNQ5 antibody (A04705-1). KCNQ5 was detected in paraffin-embedded section of rat brain tissue. Heat mediat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000185760-KCNQ5/tissue/cerebral+cortex#imid_4377000
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000185760-KCNQ5/tissue/colon#imid_4376942
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000185760-KCNQ5/tissue/liver#imid_4376943
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000185760-KCNQ5/tissue/kidney#imid_4376936
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016655; green = KCNQ5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016655; green = KCNQ5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KCNQ5 IHC & IF
🧪Run skeletal muscle first and look for staining in myocytes (HPA: High in myocytes). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); within the skeletal muscle slide, assess non-myocyte cells for background staining rather than assuming they are target negative (HPA: High in myocytes).
Positive control tissue: Skeletal muscle (Myocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KCNQ5 in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Cytoplasmic bodies (uncertain) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a rabbit IgG isotype control matched to the primary’s clonality where known, and a KCNQ5 knockout or peptide-block control (caption: rabbit anti-KCNQ5 antibody). For chromogenic detection, block endogenous peroxidase and check for endogenous biotin because the reported detection uses a biotinylated secondary and streptavidin–biotin complex with DAB (caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state a fixative (caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0 for mouse brain, but does not establish that retrieval is required for skeletal muscle (caption: mouse brain IHC and EDTA retrieval). The supplied evidence does not establish that frozen sections or IF are easier; if using IF, assess tissue autofluorescence and interpret the uncertain HPA cytoplasmic-body signal alongside UniProt’s cell-membrane annotation (HPA: cytoplasmic bodies, uncertain; UniProt Q9NR82: cell membrane).
HPA tissue IHC evidence for KCNQ5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KCNQ5 IHC Tips
Troubleshoot KCNQ5 staining in paraffin sections by checking retrieval, tissue context, subcellular pattern and controls before interpreting chromogenic signal.
What retrieval should I start with for KCNQ5 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A04705-1). The selected mouse brain image used this retrieval before incubation with 2 µg/mL primary antibody overnight at 4°C (datasheet A04705-1). If staining is weak, vary heating time in small increments while keeping section thickness, antibody concentration and detection conditions constant (standard IHC practice). Compare each condition on adjacent sections with a no-primary control, because excessive retrieval can damage morphology or increase nonspecific staining (standard IHC practice). Record neuronal staining separately from diffuse background when judging the result (HPA: medium staining in cerebral cortex neuronal cells; standard IHC practice).
Could fixation explain weak or uneven KCNQ5 staining?
Target-specific sensitivity of KCNQ5 to fixation is unknown from the supplied evidence; the selected tissue image reports paraffin embedding but does not state a fixative (datasheet A04705-1). For a fixation comparison, process matched tissue pieces with recorded fixation times, then stain adjacent sections using the same EDTA pH 8.0 retrieval and antibody conditions (datasheet A04705-1; standard IHC practice). Include a no-primary control and inspect morphology alongside signal, because delayed or uneven fixation can complicate interpretation of tissue staining (standard IHC practice). Do not assign a fixation effect solely because staining differs between tissues or subcellular compartments (standard IHC practice).
Should KCNQ5 staining be membranous or cytoplasmic?
KCNQ5 is annotated at the cell membrane and contains 6 transmembrane segments, followed by a cytoplasmic region spanning residues 347–932 (UniProt Q9NR82 subcellular location and topology). Tissue IHC has also shown cytoplasmic staining in several tissues, while the reported subcellular location of cytoplasmic bodies is uncertain (HPA: tissue profile and subcellular summary). In paraffin sections, assess whether signal follows cell outlines, cytoplasm or both, and compare that pattern with the no-primary control (standard IHC practice). Treat a new exclusively nuclear pattern cautiously and check whether it persists across adjacent sections and detection conditions (UniProt Q9NR82 subcellular location; standard IHC practice).
How could an unknown antibody epitope affect KCNQ5 staining?
KCNQ5 has 7 annotated isoforms and 6 transmembrane segments, but the supplied catalog caption does not identify the antibody epitope (UniProt Q9NR82 isoforms and topology; datasheet A04705-1). Ask for the immunogen sequence or mapped epitope before assuming every isoform is detected (standard IHC practice). An epitope in residues 347–932 would lie in the annotated cytoplasmic region, whereas extracellular loops occupy shorter intervals between membrane segments (UniProt Q9NR82 topology). If staining differs across samples, compare documented isoform coverage and retrieval conditions before attributing the difference to expression; the record also lists phosphoserines at residues 88, 447 and 831 without establishing their effect on this antibody (UniProt Q9NR82 modified residues; standard IHC practice).
How should I adapt a KCNQ5 experiment for multiplex IF?
For a separate IF experiment, pair KCNQ5 with a validated marker of the expected cell type, such as a neuronal marker when examining cerebral cortex, and verify each channel individually (HPA: medium staining in cerebral cortex neuronal cells; standard IF practice). Select fluorophores after checking tissue autofluorescence and reserve the channel with the cleanest background for a weak KCNQ5 signal (standard IF practice). The supplied antibody epitope is unmapped, so choose permeabilisation after establishing whether it faces the cytoplasm or extracellular space; KCNQ5 has both extracellular loops and extensive cytoplasmic regions (datasheet A04705-1; UniProt Q9NR82 topology). Compare permeabilised and nonpermeabilised sections with single-label and secondary-only controls before interpreting colocalisation (standard IF practice).
What should I check when DAB staining looks diffuse?
First compare the stained section with a no-primary control and inspect whether colour concentrates at tissue edges, folds or damaged areas (standard IHC practice). The selected mouse brain image used a 10% goat-serum block, biotinylated secondary antibody, streptavidin–biotin detection and DAB, so each detection step warrants a matched control when background rises (datasheet A04705-1; standard IHC practice). Use a peroxidase block and check endogenous biotin where relevant to that detection system (standard IHC practice). If controls are clean, titrate primary antibody around the documented 2 µg/mL condition and keep DAB development time identical across comparison sections (datasheet A04705-1; standard IHC practice).
How can I quantify KCNQ5 IHC across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, then apply identical imaging and DAB development settings across sections (standard IHC practice). For neuronal or myocyte staining, report the percentage of positive cells and an H-score based on intensity categories, with the denominator set to the counted cells of that population (HPA: cerebral cortex neuronal cells and skeletal muscle myocytes; standard IHC practice). If measuring stained area or positive-cell density per mm², normalise to analysable tissue area after excluding folds and damaged regions (standard IHC practice). Keep cortex and skeletal muscle results separate, and report replicate sections and control-adjusted thresholds because their reported staining levels differ (HPA: medium in cerebral cortex neuronal cells; high in skeletal muscle myocytes; standard IHC practice).
How do I distinguish specific KCNQ5 staining from artefact?
Seek a repeatable pattern in the expected cell population: neuronal cells in cerebral cortex or myocytes in skeletal muscle have reported KCNQ5 staining (HPA: medium in cerebral cortex neuronal cells; high in skeletal muscle myocytes). Compare the apparent compartment with KCNQ5’s cell-membrane annotation, while allowing for the reported cytoplasmic tissue-IHC pattern (UniProt Q9NR82 subcellular location; HPA: tissue profile). Flag staining confined to section edges, necrotic regions or folds, and investigate colour that survives no-primary or peroxidase controls (standard IHC practice). Interpret isolated strong fields cautiously: the tissue-IHC profile is rated Approved but has low consistency between antibody staining and RNA expression data (HPA: reliability description).
Boster reagents
Best KCNQ5 / Potassium voltage-gated channel subfamily KQT member 5 IHC Antibodies
A04705-1 has real paraffin-section IHC images from mouse and rat brain and human renal clear cell carcinoma (A04705-1 image captions). No IF image is supplied (catalog image inventory).
Real IHC data
Anti-KCNQ5 Antibody ®
Cat # A04705-1
A04705-1 is listed for IHC in human, mouse, and rat (catalog applications and reactivity). Its IHC images show paraffin sections of mouse and rat brain and human renal clear cell carcinoma (A04705-1 image captions).
Which to pick: For tissue IHC, choose A04705-1 for paraffin sections, starting at 2–5 μg/ml (A04705-1 image captions; datasheet: 2–5 μg/ml). For cross-species work, A04705-1 has human, mouse, and rat IHC image examples (A04705-1 image captions); the captions do not report the fixative (A04705-1 image captions). No IF/ICC-validated SKU or IF figure is supplied, and clonality is unreported (catalog applications, image inventory, and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NR82 (KCNQ5_HUMAN, Potassium voltage-gated channel subfamily KQT member 5).
- Human Protein Atlas. KCNQ5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KCNQ5 subcellular location (ICC-IF): Localized to the cytoplasmic bodies..
- Human Protein Atlas. KCNQ5 antibody validation summary (1 antibodies).
- The Changes of KCNQ5 Expression and Potassium Microenvironment in the Retina of Myopic Guinea Pigs. Frontiers in physiology 2021 — PMC8733613.
- PubMed PMID:10787416 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KCNQ5 (Potassium voltage-gated channel subfamily KQT member 5). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); cell membrane expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04705-1). Validated IHC antibody: A04705-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Skeletal muscle (Myocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04705-1); 2. Fixation: Image fixative and duration unreported (datasheet A04705-1); 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: EDTA pH 8.0 (datasheet A04705-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04705-1); 8. Primary antibody: Rabbit anti-KCNQ5, 2-5μg/ml (datasheet A04705-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04705-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Low agreement between staining and RNA (HPA tissue IHC); Isoform / epitope: 7 isoforms; check epitope isoform and membrane side (UniProt); Regulation: No staining-linked regulator reported (UniProt). Evidence: UniProt Q9NR82, HPA tissue IHC · Approved, HPA ICC-IF · cytoplasmic bodies., Datasheet A04705-1, PMC8733613. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KCNQ5/immunohistochemistry-ihc-guide.