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HVCN1 / Voltage-gated hydrogen channel 1 · IHC design guide
Design Immunohistochemistry for HVCN1
Plan chromogenic HVCN1 IHC in paraffin sections around the cytoplasmic staining seen in immune and seminiferous duct cells (HPA tissue IHC). This guide covers fixation, epitope mapping, and interpretation of cell-specific signal.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HVCN1 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 staining in immune and seminiferous duct cells (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in immune and seminiferous duct cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Lymph node+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 | Immune cells may contribute signal within other tissues (HPA tissue IHC) | |
| Regulation | Staining varies with immune-cell abundance (HPA tissue IHC) | |
| Isoform / epitope | 4 isoforms; check whether the cytoplasmic or extracellular epitope is shared (UniProt) |
Section 1
Recommended HVCN1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) appears alongside published paraffin-section methods for human tonsil (PMC3030552) and Crohn’s disease tissue (PMC8085323).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A05922); 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-HVCN1, 5 μg/mL (datasheet A05922) |
| 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 | HVCN1-positive staining in non-germinal center cells of lymph node (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in cells in seminiferous duscts and immune cells. No signal in the no-primary control. |
💡Decision noteStart with citrate HIER at pH 6.0, 95–98 °C for 20 min (page retrieval); Tris-EDTA at pH 9.0 is a published alternative (PMC3030552).
Section 2
What Is the Expected HVCN1 Staining Pattern?
HVCN1 is a four-pass membrane protein found at cell, apical and phagosomal membranes (UniProt Q96D96 topology and localization). In paraffin-section IHC, HPA describes mainly cytoplasmic staining in immune cells and cells in seminiferous ducts, with high staining in non-germinal center cells of lymph node and tonsil (HPA tissue IHC; reliability: Enhanced). Interpret the visible pattern by cell type as well as compartment.
| Distinct staining in lymph-node or tonsil non-germinal center cells, with cytoplasmic or peripheral emphasis. | This fits HPA's high cell-specific IHC signal and reported cytoplasmic pattern (HPA tissue IHC). Peripheral emphasis is compatible with membrane localization (UniProt Q96D96); chromogenic staining need not resolve an individual membrane. |
| Predominantly nuclear staining, especially when expected positive cells show little cytoplasmic signal. | An isolated nuclear pattern conflicts with the reported membrane localization and HPA cytoplasmic IHC profile (UniProt Q96D96; HPA tissue IHC). Treat it as suspect and check morphology, counterstain and a control section before scoring. |
| Strong staining in a cell population listed as undetected, or uniform staining across unrelated cell types. | That distribution does not match the cell-resolved HPA pattern (HPA tissue IHC). Cross-reactivity or endogenous chromogen activity are possibilities, especially if the no-primary control also develops color (general IHC practice). |
| Diffuse color obscures cell borders and makes positive cells hard to distinguish. | This is background rather than a reliably scoreable HVCN1 pattern. Excess detection signal or inadequate blocking or washing can contribute (general IHC practice); compare a no-primary control and intact tissue structures. |
| No staining in lymph-node or tonsil non-germinal center cells. | Absence in these high reference populations raises a sensitivity or workflow question (HPA tissue IHC). Review antibody use, retrieval, detection and section quality as general IHC checks; the payload gives no HVCN1-specific retrieval condition. |
💡Expected HVCN1 appearanceCall a result positive when the expected cells show interpretable cytoplasmic or membrane-associated color—high in lymph-node or tonsil non-germinal center cells (HPA tissue IHC; UniProt Q96D96 localization); isolated nuclear or uniform unrelated-cell staining is suspect.
| Cell population selected for review | HPA reports high staining in lymph-node and tonsil non-germinal center cells, but medium staining in appendix germinal center cells and testis elongated or late spermatids (HPA tissue IHC). Compare like cells when judging intensity. |
| Membrane topology and visible compartment | Four transmembrane segments and cytoplasmic regions support a membrane protein (UniProt Q96D96 topology). HPA nevertheless summarizes tissue IHC as cytoplasmic; avoid requiring a sharply outlined membrane in chromogenic sections (HPA tissue IHC). |
| Interpretation of low or absent staining | HPA lists lung macrophages as low and bronchial respiratory epithelial cells as not detected by tissue IHC (HPA tissue IHC). UniProt reports respiratory epithelial expression (UniProt Q96D96); those statements do not guarantee visible bronchial IHC. |
| Antibody evidence | HPA039329 has an Enhanced IHC validation status, and the tissue profile has Enhanced reliability through staining/RNA consistency (HPA antibodies; HPA tissue IHC). These ratings do not validate every cell-level signal. |
| Isoforms and epitope coverage | UniProt lists four isoforms (UniProt Q96D96). The supplied evidence does not map the IHC antibody's epitope or show isoform-specific staining, so a weak or missing signal cannot be assigned to one isoform. |
| Can the IHC pattern be applied to IF/ICC? | HPA's subcellular summary says membrane, but supplies no ICC/IF cell-line images; HPA039329 has no listed ICC validation (HPA subcellular; HPA antibodies). Use the separate IF/ICC guide for assay design. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected lymph-node or tonsil cells are unstained. | These cells are high by HPA IHC, so a missed signal is plausible; the record does not identify a target-specific fixation or retrieval effect (HPA tissue IHC). | Check section integrity and detection controls, then optimize retrieval and primary-antibody concentration as general paraffin-IHC steps. Do not infer a specific HVCN1 retrieval condition from HPA. |
| Only a nuclear deposit is visible. | Nuclear-only color conflicts with membrane localization and the reported cytoplasmic tissue profile (UniProt Q96D96; HPA tissue IHC). Counterstain or nonspecific signal may complicate reading (general IHC practice). | Inspect a no-primary control, verify focus and counterstain, and reassess whether color follows cell boundaries or cytoplasm before assigning HVCN1 positivity. |
| Color appears in nearly every cell or covers the section. | A uniform distribution is unlike HPA's cell-selective IHC profile (HPA tissue IHC). Detection background, endogenous enzyme activity or excess antibody are possible (general IHC practice). | Compare no-primary and tissue controls; review endogenous-enzyme blocking for enzyme-based detection, primary concentration and washing as general IHC checks. |
| Testis staining is weaker than lymph-node staining. | HPA rates elongated or late spermatids medium and lymph-node non-germinal center cells high (HPA tissue IHC). Equal intensity is therefore an unsuitable acceptance rule. | Score the identified cell populations separately and compare each with its HPA reference level; avoid treating the whole testis section as uniformly positive. |
| Bronchial respiratory epithelium is unstained despite reported expression. | UniProt records respiratory epithelial expression, while HPA records bronchial respiratory epithelial IHC as not detected (UniProt Q96D96; HPA tissue IHC). | Use a HPA high lymph-node or tonsil cell population to check assay performance. Do not use bronchial epithelial staining as the sole positive-control criterion. |
| A supposedly negative tissue contains a few stained cells. | HPA's negative calls identify particular cell types, such as adipocytes in adipose tissue; they are not blanket claims for every cell in a section (HPA tissue IHC). | Identify the stained cells morphologically, compare the specified HPA cell type, and inspect a no-primary control before calling the signal cross-reactive. |
Experiment evidence
IHC & IF images showing HVCN1 staining
A05922 · IHCImmunohistochemistry of HVCN1 in rat spleen tissue with HVCN1 antibody at 5 μg/mL.
A05922 · IFImmunofluorescence of HVCN1 in Rat Spleen cells with HVCN1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122986-HVCN1/tissue/cerebral+cortex#imid_10073260
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122986-HVCN1/tissue/colon#imid_10073186
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122986-HVCN1/tissue/liver#imid_10073171
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000122986-HVCN1/tissue/kidney#imid_10073172
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HVCN1 IHC & IF
🧪Run lymph node first: non-germinal center cells should stain (HPA: High). Use adipose tissue as the negative comparator, where adipocytes are not detected (HPA: Not detected); cells outside the stained compartment on the lymph-node slide should show background-level signal comparable to the no-primary control (standard IHC practice).
Positive control tissue: Lymph node (Non-germinal center cells, 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 HVCN1; derive a cell-line control from the positive tissue's cell type (Non-germinal center cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary and secondary-only controls, a host- and immunoglobulin-matched isotype control, and HVCN1-knockout tissue as a biological negative (standard IHC practice). For chromogenic staining of lymph node, block endogenous peroxidase and check the secondary-only slide for residual signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected A05922 rat-spleen IHC caption does not state a fixative (caption: fixative unreported). Retrieval dependency is also unreported, and the evidence does not establish whether frozen sections or IF/ICC are easier (supplied target/application evidence). Endogenous peroxidase can confound lymph-node chromogenic staining, so assess it with the blocked secondary-only control (standard IHC practice).
HPA tissue IHC evidence for HVCN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Rectum | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Small intestine | Enterocytes - Microvilli | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HVCN1 IHC Tips
Troubleshoot chromogenic HVCN1 staining in paraffin sections by checking retrieval, cell identity, subcellular pattern, controls, and scoring.
What retrieval conditions should I start with for HVCN1 in paraffin sections?
Use citrate pH 6.0 heat-induced antigen retrieval at 95–98 °C for 20 min in paraffin sections (page retrieval specification). Allow sections to cool in buffer, then compare staining with a matched section processed without retrieval to identify retrieval-related background (standard IHC practice). HVCN1 has 4 transmembrane segments and both cytoplasmic and extracellular loops, so retrieval may expose epitopes differently depending on antibody binding site (UniProt Q96D96 topology). If staining remains weak, test EDTA pH 9.0 on parallel sections as a fallback, holding antibody concentration and detection constant (standard IHC optimization). Score signal in the same cell populations across conditions, since stronger diffuse staining alone does not establish improved specificity (standard IHC practice).
How should I troubleshoot fixation-related loss of HVCN1 staining?
The selected HVCN1 tissue-IHC caption reports rat spleen staining at 5 µg/mL but does not state a fixative, so target-specific fixation sensitivity is unknown (A05922 caption). Record the fixative, fixation duration, tissue thickness, and processing history for each paraffin block before comparing staining intensity (standard IHC practice). If staining varies between blocks, stain adjacent sections together with the same retrieval, antibody incubation, and detection settings to reduce run-to-run variation (standard IHC practice). Compare a consistently stained control block with the affected block, including an omission control for detection background (standard IHC practice). Do not attribute a weak result specifically to HVCN1 epitope masking without a controlled fixation comparison (standard IHC practice).
Which staining compartments and cells are plausible for HVCN1?
Assess HVCN1 along cell membranes and, where appropriate, intracellular membrane-associated structures: its annotated sites include the apical membrane and phagosome membrane (UniProt Q96D96 subcellular location). Cytoplasmic staining in cells of seminiferous ducts and immune cells is also reported in tissue IHC (HPA tissue profile). For a cellular positive comparison, HPA reports high staining in lymph-node and tonsil non-germinal-center cells, with medium staining in appendix germinal-center cells (HPA tissue IHC). Interpret a broadly nuclear-only pattern cautiously because nuclear localization is absent from the supplied location annotations (UniProt Q96D96 subcellular location). Check the stained cells against tissue morphology and an antibody-omission section before assigning membrane or cytoplasmic signal to HVCN1 (standard IHC practice).
How can epitope position or isoforms change the HVCN1 IHC pattern?
HVCN1 has 4 annotated isoforms, but the supplied evidence does not map the catalog antibody’s epitope to any one isoform (UniProt Q96D96 isoforms; A05922 caption). The protein has cytoplasmic regions at residues 1–100 and 219–273, two short extracellular loops, and 4 transmembrane segments (UniProt Q96D96 topology). Consequently, apparent access to a linear epitope after retrieval depends on where the antibody binds; record the immunogen sequence if it becomes available (UniProt Q96D96 topology; standard IHC practice). Phosphothreonine 29 and phosphoserine 97 are annotated, but their effect on this antibody’s staining is untested in the supplied evidence (UniProt Q96D96 modified residues; A05922 caption). Compare localization and control tissues before calling a changed pattern isoform-specific or modification-dependent (standard IHC practice).
How should I assess HVCN1 by IF alongside a cell-type marker?
For secondary IF work, pair HVCN1 with a marker identifying the expected immune-cell population and inspect separate channels before interpreting overlap (UniProt Q96D96 tissue specificity; standard IF practice). Choose a fluorophore in a channel with low tissue autofluorescence, and include unstained and single-label controls to set exposure and assess bleed-through (standard IF practice). HVCN1 has cytoplasmic and extracellular regions, while the catalog antibody’s epitope side is unspecified; optimize permeabilisation for the mapped epitope rather than assuming it is intracellular (UniProt Q96D96 topology; A05922 caption). For a cytoplasmic epitope, compare an untreated section with a gently permeabilised section; an extracellular epitope may be accessible without permeabilisation (standard IF practice). The supplied catalog caption establishes tissue IHC staining, not IF performance, so validate IF signal independently (A05922 caption).
What should I check when HVCN1 chromogenic staining is diffuse?
First compare the stained section with an antibody-omission control and inspect whether colour follows tissue edges, folds, damaged areas, or endogenous pigment (standard IHC practice). For peroxidase-based chromogenic detection, confirm the peroxide block and review the DAB development time on matched sections; these are general workflow controls (standard IHC practice). Titrate the primary antibody around the reported 5 µg/mL rat-spleen caption concentration only as an optimization starting point, since processing and detection conditions are unreported there (A05922 caption; standard IHC practice). Shorter chromogen development or stronger blocking can clarify cell outlines if the omission control is clean but overall staining is excessive (standard IHC practice). Retain the condition that preserves plausible immune-cell or membrane-associated signal while reducing unrelated diffuse colour (HPA tissue profile; UniProt Q96D96 subcellular location).
How should I score HVCN1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HVCN1 is reported in immune cells and at several membrane sites (HPA tissue profile; UniProt Q96D96 subcellular location). For intensity, record the percentage of cells at scores 0–3 and calculate an H-score from 0–300; alternatively report percentage positive within the prespecified population (standard IHC scoring practice). For spatial studies, count positive cells per mm² of viable tissue and separately record total eligible cells or tissue area as the denominator (standard IHC quantification practice). Apply one staining threshold across the batch using the same positive reference and omission control, then review borderline cells against morphology (standard IHC practice). Report membrane-associated and cytoplasmic scores separately if both patterns occur, rather than combining unlike compartments into one value (UniProt Q96D96 subcellular location; HPA tissue profile; standard IHC practice).
How can I distinguish convincing HVCN1 staining from artefact?
A convincing result should follow recognizable cells and a plausible membrane-associated or cytoplasmic distribution, with staining in relevant immune-cell populations (UniProt Q96D96 subcellular location; HPA tissue profile). Compare lymph-node or tonsil non-germinal-center cells, reported as high, with adipose-tissue adipocytes, reported as not detected, while respecting differences between specimens (HPA tissue IHC). Treat exclusively nuclear colour, staining confined to cut edges, and deposits over necrotic areas as patterns requiring further controls before assigning HVCN1 (UniProt Q96D96 subcellular location; standard IHC practice). An antibody-omission section helps reveal endogenous enzyme activity or detection-reagent deposits in chromogenic assays (standard IHC practice). Recheck morphology, retrieval, and antibody concentration on adjacent sections before interpreting a discordant pattern as a biological change (standard IHC practice).
Boster reagents
Best HVCN1 / Voltage-gated hydrogen channel 1 IHC Antibodies
The catalog antibody has IHC data from rat spleen tissue and IF data from rat spleen cells (IHC and IF figure captions); it lists Human, Mouse and Rat reactivity (catalog: A05922).
Real IHC data
Anti-HVCN1 Antibody
Cat # A05922
A05922 is the SKU with a rendered IHC card: its figure shows rat spleen tissue stained at 5 μg/mL (IHC figure caption). The same SKU lists IHC-P and IF applications, with an IF figure from rat spleen cells at 20 μg/mL (catalog: A05922; IF figure caption).
Which to pick: For paraffin-section IHC, choose A05922 (catalog: IHC-P; IHC figure caption: rat spleen tissue); the figure caption does not report a fixative. For IF, choose A05922 (catalog: IF; IF figure caption: rat spleen cells); the payload does not establish separate ICC validation. For cross-species planning, A05922 lists Human, Mouse and Rat reactivity, while A05922-1 is a human-reactive polyclonal IHC option without a supplied IHC figure (catalog: A05922 and A05922-1).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96D96 (HVCN1_HUMAN, Voltage-gated hydrogen channel 1).
- Human Protein Atlas. HVCN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HVCN1 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. HVCN1 antibody validation summary (1 antibodies).
- HVCN1 modulates BCR signal strength via regulation of BCR-dependent generation of reactive oxygen species. Nature immunology 2010 — PMC3030552.
- Neutralization of Hv1/HVCN1 With Antibody Enhances Microglia/Macrophages Myelin Clearance by Promoting Their Migration in the Brain. Frontiers in cellular neuroscience 2021 — PMC8570284.
- Diagnostic and Predictive Value of Immune-Related Genes in Crohn's Disease. Frontiers in immunology 2021 — PMC8085323.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HVCN1 (Voltage-gated hydrogen channel 1). Expected localisation Cytoplasmic staining in immune and seminiferous duct cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A05922 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Lymph node (Non-germinal center cells); 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 A05922); 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-HVCN1, 5 μg/mL (datasheet A05922); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Immune cells may contribute signal within other tissues (HPA tissue IHC); Isoform / epitope: 4 isoforms; check whether the cytoplasmic or extracellular epitope is shared (UniProt); Regulation: Staining varies with immune-cell abundance (HPA tissue IHC). Evidence: UniProt Q96D96, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A05922, PMC3030552, PMC8085323. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HVCN1/immunohistochemistry-ihc-guide.