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TMC7 / Transmembrane channel-like protein 7 · IHC design guide
Design Immunohistochemistry for TMC7
Plan chromogenic IHC on paraffin sections using 2–5 μg/ml of the IHC-validated antibody (datasheet A14820-1). Compare staining with the reported cytoplasmic and membranous tissue pattern, while accounting for its uncertain reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TMC7 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells: cytoplasmic/membranous staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A14820-1) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Adipose tissue+3 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A14820-1) | |
| Caveat | Staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; map the epitope to extracellular or cytoplasmic regions (UniProt) |
Section 1
Recommended TMC7 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with a published pancreatic tissue microarray example (PMC6412825).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A14820-1) |
| Fixation | Image fixative and duration unreported (datasheet A14820-1); 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 A14820-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A14820-1) |
| Primary antibody | Rabbit anti-TMC7, 2-5 μg/ml (datasheet A14820-1) |
| Primary incubation | Overnight at 4 °C (datasheet A14820-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A14820-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TMC7-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and membranous expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A14820-1); the published pancreatic study does not specify retrieval conditions (PMC6412825).
Section 2
What Is the Expected TMC7 Staining Pattern?
TMC7 is a nine-pass membrane protein (UniProt Q7Z402 topology). In tissue IHC, HPA reports general cytoplasmic and membranous staining, including High staining in breast and prostate glandular cells, bronchial respiratory epithelial cells, and kidney glomerular cells (HPA tissue IHC). Treat these as provisional reference patterns: HPA rates tissue staining Uncertain because antibody staining and RNA expression have low consistency (HPA tissue IHC reliability).
| Membranous and cytoplasmic staining appears in breast or prostate glandular cells, bronchial respiratory epithelium, or kidney glomerular cells. | This resembles the reported High cell-specific IHC pattern (HPA tissue IHC). Score the relevant cells and compartments separately; agreement with HPA supports interpretation but does not independently establish specificity because tissue IHC reliability is Uncertain (HPA tissue IHC reliability). |
| The dominant signal is nuclear, with little membranous or cytoplasmic staining. | A nuclear-only pattern does not match the HPA tissue profile or UniProt membrane annotation (HPA tissue IHC; UniProt Q7Z402 subcellular location). Inspect counterstain overlap and detection controls before assigning the nuclear signal to TMC7 (general IHC practice). |
| Strong signal appears chiefly in adipocytes or skeletal myocytes rather than the expected cell population. | HPA reports TMC7 Not detected in those cell types (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogen-producing activity; compare a no-primary control and a reported positive cell population on the same run (general IHC practice; HPA tissue IHC). |
| Chromogen covers many tissue compartments without clear cell boundaries or contrast with surrounding tissue. | This does not resolve the cytoplasmic and membranous profile reported by HPA (HPA tissue IHC). Diffuse background can reflect nonspecific binding, inadequate washing, or detection-system background; inspect no-primary and detection controls before scoring cells (general IHC practice). |
| No convincing signal is seen in a section containing breast glandular or bronchial respiratory epithelial cells. | Those cells are reported High by HPA, so absent staining warrants an assay check (HPA tissue IHC). Confirm the cells are present and assess retrieval, antibody conditions, and detection with run controls; a negative result alone cannot settle expression given HPA's Uncertain tissue reliability (general IHC practice; HPA tissue IHC reliability). |
💡Expected TMC7 appearanceCall a section provisionally positive when identifiable reported cell types show clear membranous and cytoplasmic chromogen at an intensity consistent with HPA's High examples; dominant nuclear-only staining or equally strong staining in reported Not detected cell types is suspect (HPA tissue IHC; HPA tissue IHC reliability).
| Membrane topology and antibody access (UniProt Q7Z402 topology) | TMC7 has 9 transmembrane segments and both extracellular and cytoplasmic regions (UniProt Q7Z402 topology). Epitope position is not supplied, so topology alone cannot choose a retrieval condition or predict which membrane face the IHC antibody recognizes (UniProt Q7Z402 topology; evidence limit). |
| Tissue evidence and validation (HPA tissue IHC; HPA antibody HPA029465) | HPA lists Uncertain tissue IHC reliability and Uncertain IHC validation for HPA029465; its ICC validation is Approved (HPA tissue IHC; HPA antibody HPA029465). Use the tissue examples as comparisons, not a fixed pass/fail threshold (HPA tissue IHC reliability). |
| Isoforms and modifications (UniProt Q7Z402) | UniProt lists 2 isoforms and five glycosylation sites, but supplies no epitope or isoform-specific IHC pattern here (UniProt Q7Z402 isoforms and glycosylation; evidence limit). Do not infer that an absent or altered stain identifies a particular isoform or glycosylation state. |
| IF/ICC Q&A: where might TMC7 appear? (HPA subcellular ICC-IF) | HPA reports Approved vesicle localization in ICC-IF, with images from A-431, U-251MG, and U2OS (HPA subcellular ICC-IF). This complements, but does not replace, the cytoplasmic and membranous tissue IHC profile (HPA tissue IHC); IF/ICC assay conditions belong in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported positive cells are blank. | The cells may be absent from the section, or the IHC run may have failed; HPA's High examples are provisional (HPA tissue IHC reliability). | Verify cell identity on the counterstained section, then review run controls, retrieval, antibody dilution, and detection settings (general IHC practice). |
| Signal is mainly nuclear. | That compartment conflicts with the reported tissue profile and membrane annotation (HPA tissue IHC; UniProt Q7Z402 subcellular location). | Check the no-primary section and chromogen against the nuclear counterstain; only score staining that can be assigned to the reported compartments (general IHC practice; HPA tissue IHC). |
| Brown deposit appears in many cell types, including reported negative cells. | Nonspecific binding or endogenous detection activity may obscure the reported cell pattern (general IHC practice; HPA tissue IHC). | Compare no-primary and reported negative cell populations; review blocking, washes, and endogenous-peroxidase control for chromogenic IHC (general IHC practice). |
| Patchy tissue staining is hard to score. | A mixed section may contain reported positive and negative cell populations (HPA tissue IHC); section quality can also complicate scoring (general IHC practice). | Score identifiable cell types separately and compare intact tissue regions with the counterstain; exclude damaged regions from interpretation (general IHC practice). |
| A reported Not detected population stains as strongly as a reported High population. | The pattern disagrees with HPA's cell-specific observations, whose tissue reliability is Uncertain (HPA tissue IHC). | Repeat comparison with appropriate run and no-primary controls, and report the discordance rather than assigning it to TMC7 without further validation (general IHC practice; HPA tissue IHC reliability). |
| Vesicular ICC-IF signal seems inconsistent with diffuse tissue IHC staining. | HPA reports vesicles in ICC-IF and general cytoplasmic and membranous tissue IHC staining; these are observations from different applications (HPA subcellular ICC-IF; HPA tissue IHC). | Interpret each image against its own application and cell context; consult the separate IF/ICC guide for that assay (HPA subcellular ICC-IF; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing TMC7 staining
A14820-1 · IHCIHC analysis of TMC7 using anti-TMC7 antibody (A14820-1). TMC7 was detected in a paraffin-embedded section of human colorectal adenocarcinom
A14820-1 · IHCIHC analysis of TMC7 using anti-TMC7 antibody (A14820-1). TMC7 was detected in a paraffin-embedded section of human liver cancer tissue. Hea
A14820-1 · IFIF analysis of TMC7 using anti-TMC7 antibody (A14820-1). TMC7 was detected in a paraffin-embedded section of human intestinal cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000170537-TMC7/tissue/cerebral+cortex#imid_7306896
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000170537-TMC7/tissue/colon#imid_7306830
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000170537-TMC7/tissue/liver#imid_7306823
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000170537-TMC7/tissue/kidney#imid_7306824
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029465; green = TMC7, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029465; green = TMC7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TMC7 IHC & IF
🧪Run bone marrow first and look for staining in hematopoietic cells, then use adipose tissue as the negative tissue (HPA: High in bone marrow hematopoietic cells; Not detected in adipose tissue adipocytes). On the positive slide, adipocytes, if present, are candidate internal negative cells and should show only background staining (HPA: Not detected in adipocytes); confirm their identity and staining on the section.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TMC7 in A-431, U-251MG, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and matched nonimmune rabbit IgG controls, plus TMC7 knockout material if available or immunogen-peptide competition to assess antibody specificity (A14820-1 caption: rabbit primary; standard IHC control practice). Quench endogenous peroxidase before DAB detection because marrow myeloid cells can produce misleading chromogenic signal (standard IHC practice).
⚠️Feasibility: No matched source reports a TMC7-specific fixation window or fixation effect, and the selected A14820-1 paraffin-section caption does not state the fixative (A14820-1 tissue-IHC caption). That caption uses heat-mediated retrieval in EDTA at pH 8.0, providing an IHC example without establishing that this retrieval is required (A14820-1 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; endogenous peroxidase in marrow myeloid cells is a potential DAB artefact (standard IHC practice).
HPA tissue IHC evidence for TMC7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Endothelial cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TMC7 IHC Tips
Troubleshoot TMC7 chromogenic IHC in paraffin sections using the catalog antibody’s tissue image and the stated limits of its localisation evidence.
Which retrieval condition should I start with for TMC7 in paraffin sections?
Use heat-mediated retrieval in EDTA at pH 8.0 for TMC7 paraffin-section IHC (datasheet A14820-1). The catalog image used this condition before overnight incubation with 2 μg/ml antibody at 4°C (datasheet A14820-1). If staining is weak, check that sections stayed covered during heating and that the retrieval bath reached its intended temperature (standard IHC practice). Compare any subsequent retrieval adjustment on adjacent sections, keeping antibody concentration and DAB development constant (standard IHC practice). Judge improvement by cellular staining and tissue preservation together; retrieval that damages morphology makes a stronger signal difficult to interpret (standard IHC practice).
How should I troubleshoot weak TMC7 staining when fixation details are missing?
The TMC7 catalog image documents a paraffin section, but its fixative is unreported; target-specific fixation sensitivity is therefore unknown (datasheet A14820-1). Record the specimen’s actual fixative and fixation duration before comparing staining between cases (standard IHC practice). Process comparison sections together, then apply the documented EDTA pH 8.0 retrieval and 2 μg/ml antibody condition consistently (datasheet A14820-1; standard IHC practice). If one batch stains weakly, examine morphology and an established tissue control from the same run before changing retrieval (standard IHC practice). Do not assign a TMC7-specific fixation effect without a direct controlled comparison.
Should TMC7 staining appear at the cell surface or in cytoplasmic puncta?
Evaluate both membrane-associated and cytoplasmic staining: TMC7 is annotated as a membrane protein, while tissue IHC reports general cytoplasmic and membranous expression (UniProt Q7Z402; HPA tissue IHC). Approved subcellular imaging places TMC7 in vesicles, so puncta can also be biologically plausible (HPA subcellular). Its 9 predicted transmembrane segments support membrane association but do not identify which membrane generates a chromogenic signal (UniProt Q7Z402 topology). Compare the compartment and cell type across intact areas of the section, using a counterstain to define cell boundaries (standard IHC practice). Treat diffuse staining alone cautiously because the tissue IHC reliability is rated uncertain (HPA tissue IHC).
Could epitope position or alternative splicing change my TMC7 IHC pattern?
TMC7 has 2 annotated isoforms, so determine whether the catalog antibody’s documented epitope is shared before comparing specimens or methods (UniProt Q7Z402 isoforms; standard IHC practice). The protein has alternating extracellular and cytoplasmic regions around 9 predicted transmembrane segments, making epitope position relevant to accessibility (UniProt Q7Z402 topology). Five annotated glycosylation sites occur at residues 24, 84, 96, 259, and 638, but their effect on this antibody’s staining is not established (UniProt Q7Z402). Keep the validated EDTA pH 8.0 retrieval as the reference condition when investigating epitope access (datasheet A14820-1). Do not attribute a changed pattern to an isoform without isoform-specific evidence.
How can IF help assess an ambiguous TMC7 chromogenic IHC pattern?
Use IF as a separate localisation check, since this page’s documented antibody condition is paraffin-section chromogenic IHC (datasheet A14820-1). Multiplex TMC7 with a validated marker for the cell type being evaluated, then inspect whether signals occupy the same cells rather than relying on apparent overlap (standard IF practice). Select fluorophores and exposure settings against tissue autofluorescence, including an appropriate background control (standard IF practice). Plan permeabilisation around the mapped epitope: a cytoplasmic-facing region requires intracellular access, whereas an extracellular-facing epitope should be assessed with membrane integrity in mind (UniProt Q7Z402 topology; standard IF practice). HPA reports vesicular localisation in ICC/IF, which can guide comparison without establishing this catalog antibody’s IF performance (HPA subcellular).
What should I check when TMC7 DAB staining is widespread?
First inspect a no-primary control for background from the detection system, including endogenous peroxidase activity (standard IHC practice). The catalog image used a peroxidase-conjugated secondary and DAB, so apply an appropriate peroxidase block as a general chromogenic IHC step (datasheet A14820-1; standard IHC practice). Its tissue section was blocked with 10% goat serum before 2 μg/ml primary antibody overnight at 4°C (datasheet A14820-1). If controls are clean but staining is diffuse, compare a lower primary concentration and shorter DAB development on adjacent sections (standard IHC practice). Preserve interpretable membrane or vesicular cellular detail when choosing the working condition (UniProt Q7Z402; HPA subcellular; standard IHC practice).
How should I score TMC7 when positive cells and staining intensity vary? ⚠ ANSWER MARKED FOR VERIFICATION
Define the evaluated cell population and compartment before scoring, then keep those definitions fixed across cases (standard IHC practice). For mixed intensity, report an H-score from the percentages of cells at each intensity; also report the percentage of positive cells when that is the primary endpoint (standard IHC practice). If counting discrete positive cells, express density per mm² of viable, evaluable tissue rather than per whole image (standard IHC practice). Normalise comparisons to the same cell population, tissue area, staining batch, and exposure or scan settings (standard IHC practice). Record membranous and cytoplasmic patterns separately because both appear in HPA tissue IHC, whose reliability is uncertain (HPA tissue IHC).
How do I distinguish convincing TMC7 staining from section artefacts?
A convincing result follows intact cell boundaries or vesicular structures and recurs in comparable cells across the section (UniProt Q7Z402; HPA subcellular; standard IHC practice). Check cell identity carefully: HPA reports high staining in colon endothelial cells, while its overall tissue IHC reliability is uncertain (HPA tissue IHC). Staining restricted to cut edges, folds, or necrotic regions should be excluded from interpretation (standard IHC practice). Compare a no-primary control to identify endogenous enzyme or detection background before calling a positive DAB signal (standard IHC practice). A pattern outside the expected membrane-associated or vesicular compartments needs independent support before being assigned to TMC7 (UniProt Q7Z402; HPA subcellular).
Boster reagents
Best TMC7 / Transmembrane channel-like protein 7 IHC Antibodies
A14820-1 has IHC images from human colorectal adenocarcinoma and liver cancer paraffin sections, plus an IF image from a human intestinal cancer paraffin section (catalog image captions).
Real IHC data
Anti-TMC7 Antibody ®
Cat # A14820-1
A14820-1 will render with its human colorectal adenocarcinoma paraffin-section IHC image; a second IHC caption describes human liver cancer (catalog IHC image captions). Its application list includes IHC and IF, its IF caption shows human intestinal cancer paraffin tissue, and its listed reactivity is human, mouse, and rat (catalog applications, IF image caption, reactivity).
Which to pick: Choose A14820-1 for human paraffin-section chromogenic IHC: its IHC captions document EDTA pH 8.0 retrieval and DAB detection, but do not report the fixative (catalog IHC image captions). For IF, A14820-1 has a human paraffin-section image; ICC validation is not documented in the supplied payload (catalog IF image caption, applications). For mouse or rat work, A14820-1 lists reactivity with both species, while the supplied IHC and IF images show human tissue only (catalog reactivity, image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q7Z402 (TMC7_HUMAN, Transmembrane channel-like protein 7).
- Human Protein Atlas. TMC7 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TMC7 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. TMC7 antibody validation summary (1 antibodies).
- TMC7 deficiency causes acrosome biogenesis defects and male infertility in mice. eLife 2024 — PMC11398861.
- TMC7 promotes colorectal cancer metastasis via inhibition of PKN2 and regulation of hypoxia-adaptive responses. Translational oncology 2026 — PMC13400207.
- Identification of candidate diagnostic and prognostic biomarkers for pancreatic carcinoma. EBioMedicine 2019 — PMC6412825.
- PubMed PMID:12812529 — UniProt-cited evidence.
- PubMed PMID:12906855 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TMC7 (Transmembrane channel-like protein 7). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A14820-1). Validated IHC antibody: A14820-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A14820-1); 2. Fixation: Image fixative and duration unreported (datasheet A14820-1); 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 A14820-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A14820-1); 8. Primary antibody: Rabbit anti-TMC7, 2-5 μg/ml (datasheet A14820-1); 9. Primary incubation: Overnight at 4 °C (datasheet A14820-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (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 A14820-1); Caveat: Staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; map the epitope to extracellular or cytoplasmic regions (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q7Z402, HPA tissue IHC · Uncertain, HPA ICC-IF · vesicles., Datasheet A14820-1, PMC6412825. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TMC7/immunohistochemistry-ihc-guide.