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TRPV5 / Transient receptor potential cation channel subfamily V member 5 · IHC design guide
Design Immunohistochemistry for TRPV5
Plan chromogenic paraffin IHC for TRPV5 using kidney tubular staining as a reference (HPA tissue IHC). This guide covers fixation consistency, the catalog antibody’s IHC dilution of 0.5–1 μg/ml (datasheet A03218-1), and the uncertain staining reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TRPV5 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 | Observed tubular cytoplasm (HPA tissue IHC); apical membrane expected (UniProt) | |
| Staining pattern | Cytoplasmic staining in kidney tubular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A03218-1) | |
| Positive control | Kidney | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03218-1) | |
| Caveat | Presumed off-target staining; reliability uncertain (HPA tissue IHC) | |
| Regulation | WNK3 increases surface glycoform (UniProt) | |
| Isoform / epitope | 2 isoforms; check cytoplasmic vs extracellular epitope coverage (UniProt) |
Section 1
Recommended TRPV5 IHC & IF Protocols
Compare the catalog antibody’s citrate pH 6 IHC protocol (datasheet A03218-1) with published chromogenic protocols for skin tumors (PMC13299883), canine cartilage (PMC3229839), and equine cartilage (PMC3344224).
| Sample | Paraffin-embedded human pancreatic cancer tissues; fixative not specified (datasheet A03218-1) |
| Fixation | Image fixative and duration unreported (datasheet A03218-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A03218-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03218-1) |
| Primary antibody | Rabbit anti-TRPV5, 0.5-1μg/ml (datasheet A03218-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03218-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03218-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TRPV5-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a group of tissues, including in renal tubules of kidney. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet A03218-1; PMC13299883), then optimize primary dilution for the sample and antibody (PMC13299883; PMC3229839; PMC3344224).
Section 2
What Is the Expected TRPV5 Staining Pattern?
TRPV5 is a six-pass channel assigned to the apical cell membrane, with apical localisation in kidney distal tubular cells reported by similarity (UniProt Q9NQA5 topology/localisation). Kidney tubular cells show high IHC staining, although HPA describes the observed pattern as cytoplasmic and rates its tissue IHC reliability Uncertain because staining and RNA data agree poorly and presumed off-target binding was observed (HPA tissue IHC).
| Kidney tubular cells stain strongly, with signal near the apical edge and some cytoplasmic staining. | This is compatible with the predicted apical membrane location and HPA’s high tubular staining and cytoplasmic profile (UniProt Q9NQA5 localisation; HPA tissue IHC). Record membrane and cytoplasmic signal separately; the HPA profile alone does not establish that every stained structure is TRPV5 (HPA tissue IHC: Uncertain). |
| Nuclei dominate the signal, or staining is confined to a compartment away from the tubular apical region. | A dominant nuclear pattern conflicts with the annotated membrane localisation and six transmembrane segments (UniProt Q9NQA5 topology/localisation). Treat it as suspect and compare with the negative control and tissue morphology (general IHC practice). HPA’s reported cytoplasmic staining means cytoplasmic signal alone cannot be dismissed (HPA tissue IHC). |
| Strong signal appears in adipocytes or appendix glandular cells, with little staining in kidney tubules. | HPA lists those cells as not detected and kidney tubular cells as high (HPA tissue IHC). Consider cross-reactivity or detection-related activity before assigning the signal to TRPV5 (general IHC practice). HPA also reports presumed off-target binding, so its tissue pattern needs cautious interpretation (HPA tissue IHC: Uncertain). |
| DAB color spreads across the section, obscuring cell borders and tubular architecture. | Diffuse color cannot support a compartment call (general IHC practice). Compare with a no-primary control, then assess nonspecific antibody staining and endogenous detection activity if the control is colored (general IHC practice). The expected HPA observation is cellular staining in kidney tubules, not an unstructured field of color (HPA tissue IHC). |
| Kidney tubules show no convincing signal despite a readable section and detection control. | This conflicts with HPA’s high staining in kidney tubular cells (HPA tissue IHC). Check the antibody’s validated IHC-P conditions, primary-antibody concentration, retrieval conditions, and detection reagents as general troubleshooting steps (general IHC practice). Because HPA IHC reliability is Uncertain, one negative section does not settle biological absence (HPA tissue IHC). |
💡Expected TRPV5 appearanceA plausible positive is high staining in kidney tubular cells, ideally showing an apical component consistent with membrane localisation; dominant nuclear signal, diffuse color, or strong staining in HPA-listed negative cells calls for control-based review (HPA tissue IHC; UniProt Q9NQA5 localisation; general IHC practice).
| Membrane topology and compartment | TRPV5 has six transmembrane segments and an annotated apical membrane location (UniProt Q9NQA5 topology/localisation). HPA reports cytoplasmic tissue staining instead, so score the observed compartments explicitly and avoid calling cytoplasmic color proof of apical membrane localisation (HPA tissue IHC: Uncertain). |
| Tissue pattern and validation | HPA reports high staining in kidney tubular cells but rates tissue IHC Uncertain because antibody staining and RNA expression are poorly consistent and presumed off-target binding was disregarded (HPA tissue IHC). Use its positive and negative cell observations as comparison points, not definitive specificity controls. |
| Isoforms and epitope coverage | UniProt lists two TRPV5 isoforms, but the supplied record gives no antibody epitope or isoform coverage (UniProt Q9NQA5 isoforms; HPA antibody record). The isoform annotation therefore cannot explain a particular section’s staining intensity or absence of signal. |
| Detection chemistry | Endogenous activity can contribute color in enzyme-based chromogenic IHC; a no-primary control helps distinguish detection-related signal from primary-antibody-dependent staining (general IHC practice). This is a general workflow consideration, not an established TRPV5-specific effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| Tubular signal is weak or absent. | Primary-antibody concentration, retrieval, or detection may need adjustment (general IHC practice); HPA reports high kidney tubular staining but Uncertain IHC reliability (HPA tissue IHC). | Confirm that the kidney section and detection control are readable, then optimize one IHC-P variable at a time within the catalog antibody’s validated instructions (general IHC practice). Do not infer a TRPV5-specific fixation effect from these data. |
| Most cells show diffuse brown color. | Nonspecific staining or endogenous detection activity may obscure cell boundaries (general IHC practice). | Inspect a no-primary control, check blocking and washes, and reassess whether kidney tubular cells remain distinguishable (general IHC practice; HPA tissue IHC: high in kidney tubular cells). |
| Nuclear staining is stronger than tubular membrane-associated signal. | A nuclear-dominant pattern does not fit the annotated apical membrane location (UniProt Q9NQA5 localisation/topology). | Check morphology and negative controls, then score nuclear and tubular staining separately (general IHC practice). Avoid assigning nuclear color to TRPV5 without independent support. |
| Adipocytes or appendix glandular cells stain strongly. | HPA records those cell types as not detected, and its TRPV5 tissue IHC is Uncertain with presumed off-target binding observed (HPA tissue IHC). | Compare kidney tubular and negative-cell staining under the same detection conditions; investigate primary-dependent staining and detection background before interpreting the positive cells (general IHC practice). |
| A cytoplasmic signal lacks a clear apical edge. | HPA describes cytoplasmic staining, while UniProt assigns TRPV5 to the apical membrane; the supplied evidence does not resolve every cytoplasmic signal (HPA tissue IHC; UniProt Q9NQA5 localisation). | Describe the visible compartment and tubular cell type without claiming membrane-only localisation; interpret with HPA’s Uncertain reliability in view (HPA tissue IHC; general IHC practice). |
| IF/ICC: should an identical pattern be required? | HPA’s subcellular summary says membrane but lists no ICC-IF image cell lines or main location, and its antibody record provides no ICC validation entry (HPA subcellular; HPA antibody record). | Use the separate IF/ICC guide for that application. For this IHC-P result, base the call on tubular cell morphology, controls, and the stated HPA and UniProt caveats (HPA tissue IHC; UniProt Q9NQA5 localisation; general IHC practice). |
Experiment evidence
IHC & IF images showing TRPV5 staining
A03218-1 · IHCIHC analysis of TRPV5 using anti-TRPV5 antibody (A03218-1). TRPV5 was detected in paraffin-embedded section of human pancreatic cancer tissu
A03218-1 · IHCIHC analysis of TRPV5 using anti-TRPV5 antibody (A03218-1). TRPV5 was detected in paraffin-embedded section of mouse kidney tissues. Heat me
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000127412-TRPV5/tissue/cerebral+cortex#imid_18760600
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000127412-TRPV5/tissue/colon#imid_18760510
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000127412-TRPV5/tissue/liver#imid_18760519
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000127412-TRPV5/tissue/kidney#imid_18760520
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TRPV5 IHC & IF
🧪Run kidney first: cells in tubules should show TRPV5 staining, with an apical pattern expected from its reported localization (HPA: High in kidney cells in tubules; UniProt Q9NQA5: apical cell membrane). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the kidney slide, use adjacent nontubular cells as an internal background comparator and look for absence of the distinct tubular apical signal (HPA: High in kidney cells in tubules; UniProt Q9NQA5: apical cell membrane).
Positive control tissue: Kidney (Cells in 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 TRPV5; derive a cell-line control from the positive tissue's cell type (Cells in tubules) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control with clonality matched to the primary when known; and TRPV5 knockout tissue or validated immunizing-peptide competition as a biological negative (selected-SKU caption: rabbit anti-TRPV5 primary; standard IHC practice). Quench endogenous peroxidase and check endogenous biotin background in kidney when using biotin-based DAB detection (selected-SKU caption: biotinylated secondary and SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03218-1 paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The caption uses heat-mediated citrate retrieval at pH 6 for 20 min, but does not establish that retrieval is required (selected-SKU caption). The supplied evidence does not establish that frozen sections or IF/ICC are easier; kidney endogenous biotin may complicate the caption’s biotin-based detection (selected-SKU caption: biotinylated secondary and SABC/DAB; standard IHC practice).
HPA tissue IHC evidence for TRPV5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 | Cells in tubules | High | 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 → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TRPV5 IHC Tips
Use matched paraffin sections and compartment-aware controls to troubleshoot TRPV5 chromogenic IHC; interpret staining cautiously because tissue IHC reliability is uncertain (HPA: uncertain).
How should I adjust retrieval when TRPV5 staining is weak or uneven?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes on paraffin sections (datasheet A03218-1). The illustrated section was subsequently stained with 1 μg/mL primary antibody overnight at 4°C, providing a reference for the full workflow (caption A03218-1). Compare retrieval times on adjacent sections while holding antibody concentration, heating equipment, and DAB development constant; score tissue damage and background alongside signal (standard IHC practice). If staining remains weak, test another buffer or pH as a documented fallback, using kidney tubules as a cautious positive reference because HPA tissue staining has uncertain reliability (HPA: high in kidney tubular cells; uncertain tissue IHC).
Could fixation explain absent or patchy TRPV5 staining?
The illustrated paraffin section does not report its fixative, so target-specific TRPV5 fixation sensitivity is unknown (caption A03218-1: fixative not stated). Record the fixative, fixation duration, processing schedule, and section age for each specimen; compare adjacent sections processed alike before attributing a difference to fixation (standard IHC practice). Hold citrate retrieval at pH 6 for 20 minutes while assessing that variable, since those are the conditions reported for this antibody (datasheet A03218-1). Include a morphology check and a matched control tissue, and interpret weak staining only after confirming that the control completed the same detection steps (standard IHC practice).
What staining pattern should I expect in kidney tubules?
Prioritize an apical membrane pattern in kidney distal tubular cells when judging a plausible TRPV5 signal (UniProt Q9NQA5: apical membrane; distal tubular colocalization by similarity). The protein has 6 transmembrane segments, so a diffuse nuclear pattern would conflict with its annotated topology (UniProt Q9NQA5: topology). HPA reports high staining in kidney tubular cells but also describes cytoplasmic staining and rates its tissue IHC reliability uncertain because of inconsistent RNA agreement and presumed off-target binding (HPA: kidney tissue IHC). Compare apical staining across intact tubules on matched sections, documenting cytoplasmic signal separately rather than counting all brown pixels as membrane staining (standard IHC practice; UniProt Q9NQA5: apical membrane).
How can epitope location and isoforms affect this IHC result?
TRPV5 has 2 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt Q9NQA5: isoforms; caption A03218-1). Its extracellular region includes residue 358, an annotated glycosylation site, while residues 685 and 689 are annotated cytoplasmic phosphorylation sites (UniProt Q9NQA5: topology and modifications). Consequently, do not infer which modification or isoform changes staining without an epitope map and direct validation (standard antibody-validation practice). When comparing specimens, keep retrieval and antibody conditions matched, and use an independent specificity control before assigning staining to one isoform or distinguishing TRPV5 from interacting TRPV6 (UniProt Q9NQA5: TRPV6 interaction; standard IHC practice).
How should I assess TRPV5 by multiplex IF?
Treat IF as a separate assay: the supplied antibody image documents paraffin-section chromogenic IHC, and HPA lists no cell lines with ICC/IF images (caption A03218-1; HPA: subcellular record). Pair TRPV5 with an independently validated marker of kidney distal tubular cells, and assess whether signal borders the apical surface (UniProt Q9NQA5: distal tubular apical localization; standard IF practice). Choose spectrally separated fluorophores, favoring a far-red channel if tissue autofluorescence obscures shorter wavelengths, and include single-stain and secondary-only controls (standard IF practice). Because the antibody epitope is unspecified, compare minimal permeabilization for an accessible extracellular epitope with gentle permeabilization if an intracellular epitope is established; document the condition used (UniProt Q9NQA5: membrane topology; standard IF practice).
What should I check when DAB background obscures tubular staining?
The illustrated workflow used 10% goat serum, a biotinylated secondary antibody, a streptavidin–biotin complex, and DAB (caption A03218-1). Run a no-primary section to distinguish detection-system deposits from primary-dependent staining, and keep DAB development matched between sections (standard IHC practice). Include a peroxidase block in the chromogenic workflow and consider an avidin–biotin blocking comparison if the biotin-based detection system produces tissue background (standard IHC practice; caption A03218-1: biotin-based detection). Check whether brown material follows tissue edges, damaged areas, or luminal debris before assigning it to TRPV5, whose annotated location is apical membrane (standard IHC practice; UniProt Q9NQA5: apical membrane).
How should I quantify TRPV5 staining across kidney sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis region as viable kidney tubular epithelium, and record apical membrane staining separately from cytoplasmic staining because those patterns carry different interpretive weight (UniProt Q9NQA5: apical membrane; HPA: cytoplasmic tissue staining, uncertain). Report the percentage of positive tubular cells and intensity with a prespecified 0–300 H-score, or report positive tubules per mm² if tubules are the sampling unit (standard IHC scoring practice). Normalize cell-based results to the number of evaluable tubular epithelial cells, and area-based results to evaluable tissue area (standard IHC scoring practice). Apply identical thresholds, section selection, retrieval, and DAB development across specimens, with blinded review where feasible (standard IHC practice).
When is a TRPV5-positive IHC result convincing?
A reproducible apical signal in kidney distal tubules fits the annotated localization and role in calcium reabsorption (UniProt Q9NQA5: localization and function). HPA reports high staining in kidney tubular cells, but its tissue IHC reliability is uncertain and presumed off-target binding was disregarded, so staining alone is insufficient for a firm assignment (HPA: kidney tissue IHC, uncertain). Treat dominant nuclear staining, signal concentrated at cut edges or necrotic areas, and deposits persisting without primary antibody as warning signs; check endogenous enzyme activity with the appropriate detection controls (UniProt Q9NQA5: membrane topology; standard IHC practice). Confirm the compartment and cell identity with an independent antibody or orthogonal expression evidence before making a biological comparison (standard antibody-validation practice).
Boster reagents
Best TRPV5 / Transient receptor potential cation channel subfamily V member 5 IHC Antibodies
A03218-1 has paraffin-section IHC images from human pancreatic cancer and mouse and rat kidney (A03218-1 IHC captions). IF/ICC is listed, but no IF image is supplied (A03218-1 catalog).
Real IHC data
Anti-TRPV5 Antibody ®
Cat # A03218-1
A03218-1 has IHC images from paraffin sections of human pancreatic cancer and mouse and rat kidney; IHC-F and ICC are also listed applications (A03218-1 IHC captions; catalog). M03218 lists IHC and human, mouse and rat reactivity, but has no IHC image in the payload (M03218 catalog).
Which to pick: Choose A03218-1 for tissue IHC when a documented paraffin-section example matters; its captions show citrate retrieval and chromogenic detection, but do not report the fixative (A03218-1 IHC captions). For IF/ICC, A03218-1 lists ICC and a 0.5–1 μg/ml IF dilution, without an IF image (A03218-1 catalog). Both SKUs list human, mouse and rat reactivity, but A03218-1 provides IHC images in all three species; M03218 is a rabbit monoclonal IHC option without a supplied IHC image (A03218-1 IHC captions; M03218 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NQA5 (TRPV5_HUMAN, Transient receptor potential cation channel subfamily V member 5).
- Human Protein Atlas. TRPV5 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TRPV5 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. TRPV5 antibody validation summary (1 antibodies).
- Immunohistochemical Analysis of pH-Sensitive TRPV5 in Common Skin Tumors. International journal of molecular sciences 2026 — PMC13299883.
- The role of the membrane potential in chondrocyte volume regulation. Journal of cellular physiology 2011 — PMC3229839.
- Expression of transient receptor potential vanilloid channels TRPV5 and TRPV6 in retinal pigment epithelium. Molecular vision 2010 — PMC2855730.
- Expression of Transient Receptor Potential Vanilloid (TRPV) channels in different passages of articular chondrocytes. International journal of molecular sciences 2012 — PMC3344224.
- PubMed PMID:10945469 — UniProt-cited evidence.
- PubMed PMID:11549322 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TRPV5 (Transient receptor potential cation channel subfamily V member 5). Expected localisation Observed tubular cytoplasm (HPA tissue IHC); apical membrane expected (UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A03218-1). Validated IHC antibody: A03218-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human pancreatic cancer tissues; fixative not specified (datasheet A03218-1); 2. Fixation: Image fixative and duration unreported (datasheet A03218-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6 , 20 min (datasheet A03218-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03218-1); 8. Primary antibody: Rabbit anti-TRPV5, 0.5-1μg/ml (datasheet A03218-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03218-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03218-1); Caveat: Presumed off-target staining; reliability uncertain (HPA tissue IHC); Isoform / epitope: 2 isoforms; check cytoplasmic vs extracellular epitope coverage (UniProt); Regulation: WNK3 increases surface glycoform (UniProt). Evidence: UniProt Q9NQA5, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A03218-1, PMC13299883, PMC3229839. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TRPV5/immunohistochemistry-ihc-guide.