This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
TMEM87A / Golgi-pH regulating cation channel · IHC design guide
Design Immunohistochemistry for TMEM87A
Plan TMEM87A staining in paraffin sections using the granular cytoplasmic tissue pattern reported by HPA (HPA tissue IHC). This guide covers cell selection, antibody use and interpretation alongside its annotated Golgi membrane location (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TMEM87A 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 | Granular cytoplasm (HPA tissue IHC); Golgi membrane (UniProt) | |
| Staining pattern | Granular cytoplasm in glandular, neuronal and glial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A14716-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+1 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 | Brain signal depends on the cell population sampled (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; check lumenal versus cytoplasmic epitope coverage (UniProt) |
Section 1
Recommended TMEM87A IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A14716-1) is accompanied by a published TMEM87A IHC method for human lung carcinoma tissue microarrays (PMC13221295).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A14716-1) |
| Fixation | Image fixative and duration unreported (datasheet A14716-1); 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 A14716-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A14716-1) |
| Primary antibody | Rabbit anti-TMEM87A, 1-2 μg/ml (datasheet A14716-1) |
| Primary incubation | Overnight at 4 °C (datasheet A14716-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A14716-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TMEM87A-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A14716-1); the published method does not specify retrieval (PMC13221295).
Section 2
What Is the Expected TMEM87A Staining Pattern?
TMEM87A should appear mainly as granular cytoplasmic staining in positive cells on paraffin sections (HPA: tissue IHC profile). This fits its supported Golgi localization in ICC-IF and its annotation as a seven-transmembrane protein of the Golgi and cell membranes (HPA: subcellular; UniProt Q8NBN3 topology and location). HPA rates the tissue IHC profile Enhanced, citing agreement with RNA expression and similar staining by paired antibodies (HPA: tissue IHC reliability).
| Granular cytoplasmic staining in adrenal, appendix, colon or duodenal glandular cells (HPA: High in these cells). | This matches the reported tissue pattern and offers several positive-reference choices (HPA: tissue IHC). Golgi localization supports a cytoplasmic distribution, but a chromogenic section alone does not identify each granule as Golgi (HPA: subcellular; general IHC interpretation). |
| Predominantly nuclear staining, or a uniform membrane outline with little granular cytoplasm. | This differs from the reported general granular cytoplasmic IHC profile (HPA: tissue IHC). UniProt also places TMEM87A at the cell membrane, so some membrane-associated signal is plausible; an isolated membrane or nuclear pattern needs antibody and control review before assignment to TMEM87A (UniProt Q8NBN3 location; general IHC interpretation). |
| Strong staining in adipocytes or chondrocytes despite weak staining in the selected positive-reference cells. | HPA reports TMEM87A as Not detected in adipocytes and chondrocytes (HPA: tissue IHC). Treat a strong signal there as discordant with that reference; check cross-reactivity and endogenous detection activity with appropriate controls (general IHC practice). |
| Diffuse color across tissue, including spaces or cell types without a convincing granular pattern. | This is difficult to assign to the reported TMEM87A pattern (HPA: tissue IHC). Background from the detection system or nonspecific binding can obscure cellular scoring; compare the section with reagent controls and the positive-reference tissue (general IHC practice). |
| No staining in a well-preserved section containing an HPA High cell population. | An absent signal in, for example, colon glandular cells warrants a technical check before calling an experimental sample negative (HPA: High in colon glandular cells; general IHC practice). Review antibody application, detection and counterstain, then compare with a run control; the supplied sources give no TMEM87A-specific fixation sensitivity. |
💡Expected TMEM87A appearanceCall a result consistent with TMEM87A when there is clear granular cytoplasmic staining in an HPA High cell population, such as adrenal glandular cells, with interpretable tissue morphology; strong color confined to HPA Not detected adipocytes or chondrocytes, or diffuse noncellular color, is discordant or suspect background (HPA: tissue IHC; general IHC interpretation).
| Tissue and cell choice (HPA: tissue IHC) | HPA reports High staining in adrenal, appendix, colon, duodenal and epididymal glandular cells, cortical neurons, and caudate and hippocampal glia. It reports Low staining in several other listed cell populations and Not detected staining in adipocytes and chondrocytes; score the specified cell type rather than treating an entire tissue as uniformly positive or negative (HPA: tissue IHC). |
| Localization and topology (UniProt Q8NBN3; HPA: subcellular) | UniProt annotates seven transmembrane segments, a lumenal region at residues 22–225 and a cytoplasmic tail at 459–555; HPA supports mainly Golgi localization by ICC-IF. The recognized epitope is unspecified, so topology cannot predict this antibody's staining strength or whether a retrieval step is needed (UniProt Q8NBN3 topology; HPA: subcellular). |
| Isoforms and processing (UniProt Q8NBN3) | UniProt lists three isoforms, a signal peptide at residues 1–21 and a chain at 22–555. Without the antibody's epitope and isoform coverage, do not infer that every isoform will stain equally or that loss of the signal peptide explains a negative section (UniProt Q8NBN3 processing and isoforms). |
| Glycosylation (UniProt Q8NBN3) | Four glycosylation sites are annotated at residues 79, 127, 157 and 160 (UniProt Q8NBN3 glycosylation). Their existence does not establish an effect on this antibody's binding, retrieval response or tissue intensity; no such target-specific effect is supplied. |
| IF/ICC question: should fluorescence be Golgi-enriched? (HPA: subcellular) | Yes: HPA reports TMEM87A mainly at the Golgi apparatus in ICC-IF and lists A-431, U-251MG and U2OS among imaged cell lines (HPA: subcellular). That supports a Golgi-enriched expectation for IF/ICC, while the tissue IHC observations above remain the reference for scoring paraffin sections (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-reference glandular cells have no signal. | The section may have a detection or antibody-application problem; HPA reports High staining in these selected cells (HPA: tissue IHC; general IHC practice). | Inspect morphology and the run's positive control, then check reagent order, antibody application and detection development before interpreting experimental negatives (general IHC practice). |
| Only weak staining appears in lung alveolar type I cells or liver cholangiocytes. | These cells are reported as Low, so weak staining alone is not discordant (HPA: tissue IHC). | Compare the run with a listed High cell population and score intensity within the named cell type, using consistent viewing conditions (HPA: tissue IHC; general IHC practice). |
| Strong color appears in adipocytes or chondrocytes. | This conflicts with their Not detected HPA observations and may reflect nonspecific binding or detection activity (HPA: tissue IHC; general IHC practice). | Compare serial or matched sections using an appropriate reagent control; review whether color follows cells, tissue edges or deposits before assigning it to TMEM87A (general IHC practice). |
| Granular signal is hidden by diffuse background. | Nonspecific antibody binding or the detection system may reduce contrast (general IHC practice). | Review blocking, washes and chromogen development against the run controls; retain the reported granular cytoplasmic pattern as the interpretive reference (general IHC practice; HPA: tissue IHC). |
| Staining is predominantly nuclear or exclusively outlines the plasma membrane. | That pattern does not match HPA's general granular cytoplasmic IHC profile, although UniProt also annotates cell-membrane localization (HPA: tissue IHC; UniProt Q8NBN3 location). | Check the positive-reference tissue and reagent controls; record any membrane signal separately from the granular cytoplasmic pattern rather than assuming that compartment alone validates the section (general IHC practice). |
| Two antibodies give different tissue patterns. | HPA's Enhanced rating reflects agreement between paired antibodies and RNA expression for its assessed profile; it does not guarantee agreement in every new run (HPA: tissue IHC reliability). | Compare the same named cell populations and controls across matched sections, and investigate discordant compartments or background before choosing a pattern for interpretation (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing TMEM87A staining
A14716-1 · IHCIHC analysis of TMEM87A using anti-TMEM87A antibody (A14716-1). TMEM87A was detected in a paraffin-embedded section of human colon adenocarc
A14716-1 · IHCIHC analysis of TMEM87A using anti-TMEM87A antibody (A14716-1). TMEM87A was detected in a paraffin-embedded section of human liver cancer ti
A14716-1 · IFIF analysis of TMEM87A using anti-TMEM87A antibody (A14716-1). TMEM87A was detected in an immunocytochemical section of U87 cells. Enzyme an
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000103978-TMEM87A/tissue/cerebral+cortex#imid_4891180
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000103978-TMEM87A/tissue/colon#imid_4891090
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000103978-TMEM87A/tissue/liver#imid_4888611
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000103978-TMEM87A/tissue/kidney#imid_4891092
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018104; green = TMEM87A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018104; green = TMEM87A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TMEM87A IHC & IF
🧪Run colon first and expect staining in glandular cells (HPA: High in colon glandular cells); run adipose tissue as the negative comparison and expect adipocytes to be unstained (HPA: Not detected in adipocytes). On the colon slide, use non-glandular cells as a background comparator and expect little signal relative to glandular cells, while treating their target status as unverified (HPA: High is reported for colon glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TMEM87A in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a rabbit IgG isotype control matched to the primary antibody’s clonality; and, if available, TMEM87A-knockout tissue or a validated peptide-block control (selected caption: rabbit anti-TMEM87A antibody). Quench endogenous peroxidase and inspect colon glands, luminal material and inflammatory cells for nonspecific DAB signal (selected caption: HRP/DAB detection in colon adenocarcinoma).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A14716-1 paraffin-section caption does not state the fixative (selected caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 preceded staining, so assess retrieval performance when transferring the method to other specimens (selected caption: heat-mediated EDTA retrieval); the evidence does not establish whether frozen sections or IF are easier. IF/ICC can provide a Golgi-localization cross-check (HPA: supported Golgi localization), while colon IHC requires attention to nonspecific DAB signal in luminal material and inflammatory cells (selected caption: colon adenocarcinoma, HRP/DAB detection).
HPA tissue IHC evidence for TMEM87A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Paired antibodies with high similarity supports the protein expression profile.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced TMEM87A IHC Tips
Troubleshoot chromogenic TMEM87A IHC in paraffin sections using the catalog antibody’s tissue example, protein topology, and reported staining patterns.
Which retrieval condition should I start with for TMEM87A paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A14716-1). The catalog antibody detected TMEM87A in a paraffin-embedded human colon adenocarcinoma section after this retrieval condition (datasheet A14716-1). Keep heating, cooling, and section thickness consistent across the comparison set, and record them alongside staining results (standard IHC practice). If signal is weak, first check retrieval uniformity and whether the primary antibody reaches the tissue evenly (standard IHC practice). Judge any retrieval adjustment against a concurrently stained positive section, because stronger DAB alone does not establish specific TMEM87A detection (standard IHC practice).
How should I investigate fixation-related loss of TMEM87A staining?
Target-specific sensitivity of TMEM87A staining to fixation is unknown from the supplied evidence; the catalog tissue caption identifies a paraffin-embedded section but does not state its fixative (datasheet A14716-1). Record fixative, fixation duration, tissue thickness, and processing history for each block before comparing staining (standard IHC practice). Use matched sections where possible, with the same EDTA pH 8.0 retrieval and detection workflow (datasheet A14716-1; standard IHC practice). If one block is weak, assess morphology and staining of an appropriate control alongside it before attributing the difference to fixation (standard IHC practice). Do not infer fixation sensitivity from expression patterns or predicted membrane topology (HPA tissue IHC; UniProt Q8NBN3 topology).
Where should convincing TMEM87A staining appear in a tissue section?
Expect predominantly granular cytoplasmic staining at tissue-section resolution, consistent with the reported Golgi localization of TMEM87A (HPA tissue IHC; HPA subcellular). UniProt also annotates the Golgi apparatus membrane, cell membrane, and ruffles, so a membrane-associated component is biologically plausible but should be judged against the dominant pattern (UniProt Q8NBN3 subcellular). Compare the distribution within individual cells, rather than scoring all diffuse cytoplasmic DAB as equivalent (standard IHC practice). Include a section with expected glandular-cell staining, such as colon, in the same run (HPA: High in colon glandular cells). Revisit strong nuclear-only or uniformly extracellular signal with controls before calling it TMEM87A (HPA subcellular; standard IHC practice).
How can epitope position and isoforms explain discordant TMEM87A staining?
TMEM87A has 3 annotated isoforms and 7 transmembrane segments, but the supplied catalog caption does not identify the antibody epitope or isoform coverage (UniProt Q8NBN3 isoforms and topology; datasheet A14716-1). The annotated lumenal region spans residues 22–225, while the C-terminal cytoplasmic region spans 459–555 (UniProt Q8NBN3 topology). Glycosylation sites at 79, 127, 157, and 160 make epitope accessibility a reasonable question, without establishing an effect on this antibody (UniProt Q8NBN3 glycosylation). Check the immunogen or epitope specification before interpreting discordance as isoform biology (standard IHC practice). Compare matched sections and controls under identical retrieval conditions while that uncertainty remains (standard IHC practice).
How should I plan IF follow-up to a chromogenic TMEM87A result?
Treat IF/ICC as a separate assay: the supplied catalog example establishes paraffin-section chromogenic detection, while HPA reports Golgi localization from ICC/IF images (datasheet A14716-1; HPA subcellular). Multiplex TMEM87A with a marker for the expected cell type in the specimen, then evaluate its intracellular pattern within marker-positive cells (HPA tissue IHC; standard IF practice). Choose fluorophores after checking tissue autofluorescence in unstained and secondary-only controls, favoring a cleaner spectral channel for the weaker signal (standard IF practice). Plan permeabilisation around the antibody’s epitope side of the membrane; TMEM87A has both lumenal and cytoplasmic regions, and this antibody’s epitope is unspecified (UniProt Q8NBN3 topology; datasheet A14716-1). Optimise IF fixation and permeabilisation with IF controls rather than transferring the paraffin-section settings (standard IF practice).
What should I change when TMEM87A DAB staining is widespread?
First compare the stained section with a no-primary control and inspect whether pigment follows tissue structures, edges, or damaged areas (standard IHC practice). For peroxidase-based DAB detection, verify an endogenous peroxidase block and a suitable blocking step before changing antibody concentration (standard IHC practice). The catalog tissue example used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary (datasheet A14716-1). Check washes and secondary-antibody background under the same detection conditions (standard IHC practice). A granular cytoplasmic pattern is the reported tissue profile, so uniform nuclear or extracellular deposition needs closer scrutiny (HPA tissue IHC; standard IHC practice).
How should I score TMEM87A IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, using granular cytoplasmic signal as the primary tissue pattern (HPA tissue IHC; standard IHC practice). An H-score can combine the percentage of cells at each intensity level into a 0–300 summary; also retain the underlying percentage-positive and intensity data (standard IHC practice). If the question concerns spatial abundance, report positive-cell density per mm² of viable tissue or of the defined compartment (standard IHC practice). Normalize comparisons to the same cell population, viable area, staining run, and image-analysis thresholds (standard IHC practice). Record edge and necrotic regions separately so their DAB signal does not inflate the score (standard IHC practice).
When is an apparent TMEM87A-positive cell likely an artefact?
A convincing result should fit the expected cellular distribution and predominantly granular cytoplasmic pattern, with Golgi localization providing a subcellular reference (HPA tissue IHC; HPA subcellular). HPA reports high staining in colon glandular cells and no detected staining in adipocytes, which can guide comparison without making either result an absolute specimen-level rule (HPA tissue IHC). Be cautious with staining restricted to section edges, necrotic regions, or areas showing endogenous enzyme signal in controls (standard IHC practice). Strong nuclear-only staining conflicts with the reported main Golgi location and warrants review (HPA subcellular; standard IHC practice). Confirm an unexpected cell or compartment call using morphology, matched controls, and reproducibility across sections (standard IHC practice).
Boster reagents
Best TMEM87A / Golgi-pH regulating cation channel IHC Antibodies
A14716-1 has IHC images from paraffin sections of human colon adenocarcinoma, human liver cancer, mouse brain and rat brain, plus an IF/ICC image from U87 cells (catalog image captions).
Real IHC data
Anti-TMEM87A Antibody ®
Cat # A14716-1
A14716-1 has IHC images from paraffin sections of human colon adenocarcinoma and liver cancer, and mouse and rat brain (catalog IHC captions). A14716-1 also has an IF/ICC image from U87 cells (catalog IF caption).
Which to pick: Choose A14716-1 for tissue IHC because its images document paraffin sections with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC captions); the fixative is unreported (catalog IHC captions). Choose A14716-1 for IF/ICC in U87 cells because its IF image documents staining at 5 μg/ml (catalog IF caption). For cross-species IHC, A14716-1 lists human, mouse and rat reactivity and has paraffin-section images for all three (catalog applications/reactivity; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NBN3 (GPHCC_HUMAN, Golgi-pH regulating cation channel).
- Human Protein Atlas. TMEM87A tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. TMEM87A subcellular location (ICC-IF): Mainly localized to the Golgi apparatus..
- Human Protein Atlas. TMEM87A antibody validation summary (2 antibodies).
- TMEM87A suppresses ferroptosis and increases cancer immunotherapy resistance by maintaining the Golgi apparatus pH homeostasis. Nature cancer 2026 — PMC13221295.
- Targeting the Calcineurin Homologous Protein 1 (CHP1)-Transmembrane Protein 87A (TMEM87A) mechanosensing complex: a druggable vulnerability in metastatic ovarian cancer. Molecular biomedicine 2026 — PMC13247001.
- GolpHCat (TMEM87A), a unique voltage-dependent cation channel in Golgi apparatus, contributes to Golgi-pH maintenance and hippocampus-dependent memory. Nature communications 2024 — PMC11239671.
- PubMed PMID:16303743 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TMEM87A (Golgi-pH regulating cation channel). Expected localisation Granular cytoplasm (HPA tissue IHC); Golgi membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A14716-1). Validated IHC antibody: A14716-1 at 1-2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A14716-1); 2. Fixation: Image fixative and duration unreported (datasheet A14716-1); 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 A14716-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A14716-1); 8. Primary antibody: Rabbit anti-TMEM87A, 1-2 μg/ml (datasheet A14716-1); 9. Primary incubation: Overnight at 4 °C (datasheet A14716-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Brain signal depends on the cell population sampled (HPA tissue IHC); Isoform / epitope: 3 isoforms; check lumenal versus cytoplasmic epitope coverage (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q8NBN3, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the Golgi apparatus., Datasheet A14716-1, PMC13221295. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TMEM87A/immunohistochemistry-ihc-guide.