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AQP6 / Aquaporin-6 · IHC design guide
Design Immunohistochemistry for AQP6
Plan chromogenic IHC on paraffin sections using the catalog antibody at 0.5–1 μg/ml (datasheets: A10607, PA2092). Use distal renal tubules as a positive reference, and assess cytoplasmic staining in light of reported off-target binding (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AQP6 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 renal tubule staining (HPA tissue IHC); vesicle membrane location (UniProt) | |
| Staining pattern | Selective cytoplasmic staining in distal renal tubules (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PA2092) | |
| Caveat | Presumed off-target staining has been observed (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No isoforms or processing; epitope position may matter (UniProt) |
Section 1
Recommended AQP6 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with published AQP6 paraffin-section IHC protocols (PMC2810475; PMC12399404).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA2092); 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-AQP6, 0.5-1μg/ml (datasheet PA2092) |
| 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 | AQP6-positive staining in distal tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in renal tubules. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval rule); the published ovarian IHC method does not specify retrieval conditions (PMC12399404).
Section 2
What Is the Expected AQP6 Staining Pattern?
AQP6 is a six-pass protein of cytoplasmic vesicle membranes (UniProt Q13520 topology and subcellular location). In paraffin-section IHC, expect selective cytoplasmic staining in renal distal tubules, where HPA reports high expression (HPA: kidney distal tubules, High). HPA rates its tissue IHC evidence Enhanced, while noting medium agreement with RNA data and presumed off-target staining that was disregarded (HPA: reliability).
| Strong, selective cytoplasmic staining in renal distal tubules. | This matches the reported positive pattern (HPA: distal tubules, High; selective cytoplasmic expression). Vesicle-associated staining is consistent with AQP6 location (UniProt Q13520: cytoplasmic vesicle membrane). |
| Predominantly nuclear staining, with little cytoplasmic distal-tubule signal. | This does not match the reported compartment (HPA: selective cytoplasmic expression; UniProt Q13520: cytoplasmic vesicle membrane). Review staining specificity and slide interpretation before scoring it as AQP6. |
| Strong staining in cells outside the expected renal tubules. | Treat this cautiously: HPA reports selective renal-tubule expression and notes presumed off-target staining (HPA: tissue profile and reliability). Cross-reactivity or endogenous detection activity are possible explanations (general IHC practice). |
| Diffuse staining across tissue and background, without a selective tubular pattern. | This is hard to interpret as AQP6 because the reported pattern is selective (HPA: tissue profile). Excess antibody, inadequate blocking, or incomplete washing can raise background (general IHC practice). |
| No staining in renal distal tubules. | The expected positive compartment is missing (HPA: distal tubules, High). Check tissue preservation, retrieval, antibody conditions, and detection controls as general IHC troubleshooting steps; the supplied sources do not establish AQP6-specific fixation sensitivity. |
💡Expected AQP6 appearanceCall a result positive when renal distal tubules show high, selective cytoplasmic staining (HPA: distal tubules, High; selective cytoplasmic expression); isolated nuclear or widespread nonselective staining does not match that pattern (HPA: tissue profile; UniProt Q13520: cytoplasmic vesicle membrane).
| Cell and tissue distribution | HPA reports kidney-enriched RNA and high staining in distal tubules (HPA: RNA specificity; distal tubules, High). Its sampled adipocytes and several other listed cell types were not detected (HPA: tissue IHC); use the exact sampled cell type when comparing controls. |
| Subcellular interpretation | AQP6 has six transmembrane segments and is assigned to cytoplasmic vesicle membranes (UniProt Q13520 topology and subcellular location). The tissue IHC profile is cytoplasmic (HPA: tissue profile); it does not resolve individual vesicles. |
| Evidence strength | The listed antibody, HPA015278, has Enhanced IHC status (HPA: antibody validation). HPA also reports medium staining–RNA consistency and disregarded presumed off-target binding (HPA: reliability); assess unexpected staining against the documented tissue pattern. |
| IF/ICC: should the same pattern be assumed? | A membrane summary is listed, but no ICC-IF image cell lines or main location are available (HPA: subcellular). UniProt assigns AQP6 to cytoplasmic vesicle membranes (UniProt Q13520); the supplied IF record cannot establish an observed cell-line staining pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Distal tubules are unstained in a kidney section. | The expected positive signal is absent (HPA: distal tubules, High). Tissue or detection failure is possible (general IHC practice); AQP6-specific fixation sensitivity is unreported in the supplied sources. | Confirm the distal-tubule region and run suitable detection controls; review the antibody's validated IHC conditions before adjusting retrieval or dilution (general IHC practice). |
| Signal appears mainly nuclear. | Nuclear localisation conflicts with the reported cytoplasmic tissue pattern and vesicle-membrane assignment (HPA: tissue profile; UniProt Q13520: subcellular location). | Check whether the signal persists in the expected distal-tubule cytoplasm, then review control slides and staining specificity before scoring it (general IHC practice). |
| Many unrelated cells stain strongly. | The distribution is broader than HPA's selective renal-tubule profile; HPA also notes presumed off-target binding (HPA: tissue profile and reliability). Endogenous detection activity is another possibility (general IHC practice). | Compare with a no-primary control and inspect the pattern in distal tubules; address endogenous detection activity if the control shows signal (general IHC practice). |
| The whole section has diffuse chromogen background. | A nonselective field obscures the reported tubular pattern (HPA: tissue profile). Antibody concentration, blocking, washing, or detection conditions may contribute (general IHC practice). | Check no-primary background and the validated antibody dilution; adjust blocking, washing, or detection conditions one at a time (general IHC practice). |
| A proposed negative-control tissue shows staining. | HPA's “Not detected” calls apply to specified sampled cells, including adipocytes in adipose tissue and glandular cells in adrenal gland (HPA: tissue IHC); they do not guarantee every cell in those sections is negative. | Identify the stained cell type before interpreting the control, compare it with HPA's cell-specific call, and review no-primary staining if needed (HPA: tissue IHC; general IHC practice). |
| Fluorescence appears at a cell boundary in an IF/ICC image. | The HPA subcellular summary says membrane but provides no ICC-IF image cell lines or main location (HPA: subcellular). That record cannot validate a specific IF pattern. | Treat the image as an IF observation requiring its own controls; use the vesicle-membrane assignment as context without claiming IF validation from the supplied record (UniProt Q13520: subcellular location; HPA: subcellular). |
Experiment evidence
IHC & IF images showing AQP6 staining
PA2092 · IHCAnti-Aquaporin 6 antibody, PA2092, IHC(P) IHC(P): Human Placenta Tissue
PA2092 · IHCAnti-Aquaporin 6 antibody, PA2092, IHC(P) IHC(P): Human Lung Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000086159-AQP6/tissue/cerebral+cortex#imid_17534095
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000086159-AQP6/tissue/colon#imid_17534013
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000086159-AQP6/tissue/liver#imid_17534022
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000086159-AQP6/tissue/kidney#imid_17534023
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AQP6 IHC & IF
🧪Run kidney first: distal tubule cells should stain strongly (HPA: High in kidney distal tubules). Use adipose tissue as a negative tissue, with adipocytes showing no specific staining (HPA: Not detected in adipocytes); on the kidney slide, use only independently verified AQP6-negative cells as internal negatives, which should show background staining.
Positive control tissue: Kidney (Distal tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for AQP6; derive a cell-line control from the positive tissue's cell type (Distal tubules) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and immunoglobulin class, and AQP6-knockout tissue or cognate-peptide competition if available (standard IHC practice). For chromogenic kidney sections, quench endogenous peroxidase and check for endogenous biotin when using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of antigen retrieval are unreported in the supplied evidence; optimise retrieval empirically for the IHC-validated antibody. Frozen sections or IF cannot be judged easier from the supplied evidence, and the selected PA2092 tissue-IHC caption does not report a fixative (PA2092 caption: fixative not stated). Assess kidney background from endogenous peroxidase and, if relevant to the detection system, endogenous biotin (standard IHC practice).
HPA tissue IHC evidence for AQP6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Kidney | Distal tubules | High | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AQP6 IHC Tips
Troubleshoot AQP6 staining in paraffin sections against its expected intracellular localisation and renal tubule expression (UniProt Q13520; HPA tissue IHC).
Which retrieval conditions should I try first for AQP6 IHC?
Use citrate at pH 6.0 for heat induced antigen retrieval at 95–98 °C for 20 min (page retrieval rule). AQP6 has 6 transmembrane segments and a cytoplasmic C terminus, but its antibody epitope is unspecified, so hold retrieval time and cooling conditions constant while comparing sections (UniProt Q13520 topology; catalog caption; standard IHC practice). If staining remains weak, test an alternative retrieval condition on adjacent sections and check whether background rises alongside signal (standard IHC practice). Judge the result in distal renal tubules, where strong staining is reported, using the same detection settings across conditions (HPA tissue IHC; standard IHC practice).
How should I assess whether fixation is reducing AQP6 staining?
Target specific fixation sensitivity is unknown: the PA2092 tissue caption reports paraffin IHC in human placenta but gives no fixative, fixation duration, or comparative fixation result (PA2092 tissue IHC caption). For a controlled comparison, record the fixative and duration for each specimen, then stain sections together with identical retrieval and detection conditions (standard IHC practice). Include a kidney section with distal tubules as a positive reference, while keeping tissue processing and section thickness comparable (HPA tissue IHC; standard IHC practice). If staining changes, interpret that result as a condition dependent observation until matched specimens separate fixation from retrieval and detection effects (standard IHC practice).
Where should convincing AQP6 chromogenic staining appear?
Look for a selective cytoplasmic pattern in renal tubules, with distal tubules providing the reported strong positive reference (HPA tissue IHC). AQP6 is assigned to cytoplasmic vesicle membranes, so a vesicular or granular cytoplasmic pattern is compatible with its annotated compartment (UniProt Q13520 subcellular location; standard IHC interpretation). Compare the distribution within the same section against unstained neighboring structures and an appropriate negative control, using matched counterstaining and exposure during image review (standard IHC practice). Diffuse staining across unrelated cell types should prompt a specificity check, particularly because the tissue atlas reports presumed off target binding that was disregarded (HPA tissue IHC).
How does AQP6 topology affect epitope troubleshooting?
The record annotates one 282 amino acid chain, no isoforms, and no glycosylation or modified residue sites; these annotations do not identify the catalog antibody’s epitope (UniProt Q13520 record; PA2092 tissue IHC caption). AQP6 has 6 membrane spans, a cytoplasmic N terminus, and a cytoplasmic C terminus, while several short loops face outward (UniProt Q13520 topology). Ask for the immunogen or mapped epitope before attributing weak staining to a particular loop or terminus (standard IHC practice). When comparing antibodies, require concordant staining in distal renal tubules and inspect discordant cytoplasmic patterns against negative tissues (HPA tissue IHC; standard IHC practice).
How can I check AQP6 by multiplex immunofluorescence?
Pair AQP6 with a validated distal tubule marker to test whether signal falls in the expected cell population, and include single stain controls for channel bleed through (HPA tissue IHC; standard IF practice). Choose a far red fluorophore when shorter wavelength tissue autofluorescence obscures weak signal, then inspect unstained and secondary only controls at matched settings (standard IF practice). If the antibody recognizes a cytoplasmic terminus, titrate mild permeabilisation to expose intracellular vesicles; if it recognizes an outward facing loop, compare permeabilised and unpermeabilised sections (UniProt Q13520 topology; standard IF practice). The supplied HPA record lists no ICC/IF images, so establish IF specificity independently of its tissue IHC pattern (HPA subcellular record; standard IF practice).
What should I change when brown staining appears broadly?
Compare the distribution with a kidney positive control and a tissue expected to lack detectable AQP6, such as adipose tissue adipocytes, before changing the primary antibody concentration (HPA tissue IHC). Run a no primary control, check endogenous peroxidase blocking, and shorten chromogen development if background develops without primary antibody (standard chromogenic IHC practice). If background tracks tissue edges or damaged areas, review section adhesion, washing, and drying during processing (standard IHC practice). Broad signal across unrelated cells warrants caution because the tissue atlas describes presumed off target binding despite its enhanced reliability designation (HPA tissue IHC).
How should I quantify AQP6 staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define distal renal tubules as the primary scoring compartment and apply the same region selection rules to every section (HPA tissue IHC; standard IHC practice). An H score combines the percentage of cells at intensity grades 0–3, producing a possible range of 0–300; also report the percentage of positive distal tubule cells (standard IHC scoring practice). Normalize counts to the number of evaluable distal tubule cells or stained area to the measured distal tubule area, and record excluded damaged regions (standard IHC practice). Keep retrieval, chromogen development, imaging, and positivity thresholds fixed across the comparison, with a shared kidney control on each staining run (HPA tissue IHC; standard IHC practice).
How do I distinguish genuine AQP6 staining from artefact?
Give greatest weight to selective cytoplasmic or vesicular staining in renal distal tubules, matching the reported tissue pattern and annotated intracellular membrane location (HPA tissue IHC; UniProt Q13520 subcellular location). Question strong signal in unrelated cells or an exclusively nuclear pattern, and compare it with negative tissues and a no primary control (HPA tissue IHC; standard IHC practice). Edge accentuation, necrotic areas, and residual endogenous peroxidase can produce misleading brown deposits, so exclude affected regions and inspect the peroxidase control (standard chromogenic IHC practice). Treat the PA2092 placenta image as evidence of a paraffin IHC application, not proof of fixation conditions or tissue wide specificity (PA2092 tissue IHC caption).
Boster reagents
Best AQP6 / Aquaporin-6 IHC Antibodies
The catalog includes human paraffin-section IHC images for AQP6 with PA2092 (PA2092 image captions: human placenta and lung cancer tissue); no IF images are supplied (catalog: IF image alts empty).
Real IHC data
Anti-Aquaporin 6/AQP6 Antibody ®
Cat # PA2092
PA2092 is the only SKU that will render; its IHC-P captions show human placenta and human lung cancer tissue (PA2092 image captions). Its application list includes IHC and WB, with human reactivity (PA2092 catalog entry).
Which to pick: Choose PA2092 for human paraffin-section IHC because its own captions document staining in human placenta and lung cancer tissue (PA2092 image captions); the fixative is unreported (PA2092 image captions). For cross-species IHC, consider A10607, which lists human, mouse and rat reactivity, though it has no supplied IHC image (A10607 catalog entry). Neither SKU lists IF/ICC as an application or supplies an IF image, so neither has documented IF/ICC validation here (A10607 and PA2092 catalog entries).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13520 (AQP6_HUMAN, Aquaporin-6).
- Human Protein Atlas. AQP6 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. AQP6 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. AQP6 antibody validation summary (1 antibodies).
- Aquaporin-6 is expressed along the rat gastrointestinal tract and upregulated by feeding in the small intestine. BMC physiology 2009 — PMC2765416.
- Aquaporin-6 expression in the cochlear sensory epithelium is downregulated by salicylates. Journal of biomedicine & biotechnology 2010 — PMC2810475.
- Aquaporin 2 is differentially expressed in granulosa cells of various stages of human follicles and is regulated by luteinizing hormone. Frontiers in cell and developmental biology 2025 — PMC12399404.
- Expression of adrenergic and cholinergic receptors in murine renal intercalated cells. The Journal of veterinary medical science 2014 — PMC4272982.
- PubMed PMID:8812490 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AQP6 (Aquaporin-6). Expected localisation Cytoplasmic renal tubule staining (HPA tissue IHC); vesicle membrane location (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: PA2092 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Distal tubules); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA2092); 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-AQP6, 0.5-1μg/ml (datasheet PA2092); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PA2092); Caveat: Presumed off-target staining has been observed (HPA tissue IHC); Isoform / epitope: No isoforms or processing; epitope position may matter (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt Q13520, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet PA2092, PMC2810475, PMC12399404. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AQP6/immunohistochemistry-ihc-guide.