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TMEM175 / Endosomal/lysosomal proton channel TMEM175 · IHC design guide
Design Immunohistochemistry for TMEM175
Plan paraffin-section TMEM175 IHC using caudate glia or testis Sertoli cells as high-staining references (HPA tissue IHC). Assess the reported nuclear and nuclear-rim pattern in light of TMEM175’s annotated endosomal and lysosomal membrane location (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TMEM175 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 | Nuclear/rim in tissue (HPA tissue IHC); endolysosomal membrane (UniProt) | |
| Staining pattern | General nuclear and nuclear rim staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A11712-1) | |
| Positive control | Caudate+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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Nuclear tissue staining differs from the annotated membrane location (HPA tissue IHC; UniProt) | |
| Regulation | Staining-linked expression regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; map the epitope to lumenal or cytoplasmic regions (UniProt) |
Section 1
Recommended TMEM175 IHC & IF Protocols
The catalog antibody protocol is paired with one published chromogenic IHC method for TMEM175 (PMC13018187).
| Sample | Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A11712-1) |
| Fixation | Image fixative and duration unreported (datasheet A11712-1); 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 A11712-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A11712-1) |
| Primary antibody | Rabbit anti-TMEM175, 2-5 μg/ml (datasheet A11712-1) |
| Primary incubation | Overnight at 4 °C (datasheet A11712-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A11712-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TMEM175-positive staining in glial cells of caudate (HPA tissue IHC: High). HPA tissue profile: General nuclear and nuclear membranous expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A11712-1); the published method does not specify retrieval conditions (PMC13018187).
Section 2
What Is the Expected TMEM175 Staining Pattern?
TMEM175 is a 12-pass endosome/lysosome membrane protein (UniProt Q9BSA9 topology and subcellular location). In paraffin IHC, HPA reports general nuclear and nuclear-membranous staining, with high signal in caudate glial cells and testicular Sertoli cells (HPA tissue IHC). HPA rates the tissue IHC pattern Approved, with medium consistency against RNA data and external verification pending (HPA tissue IHC). Interpret the observed nuclear pattern in light of its difference from the UniProt location.
| Prominent nuclear and nuclear-rim staining in caudate glial cells or testicular Sertoli cells (HPA tissue IHC). | This matches HPA's reported compartment and High staining in those cells (HPA tissue IHC). Record the cell type, intensity and compartment together. The pattern is an IHC observation; UniProt places TMEM175 on endosome and lysosome membranes (UniProt Q9BSA9). |
| Only coarse cytoplasmic deposits, diffuse cytosolic fill or extracellular chromogen in a reference-positive region. | These appearances do not match HPA's reported nuclear and nuclear-membranous IHC profile (HPA tissue IHC). Endolysosomal puncta remain biologically plausible from the UniProt location, so an alternate compartment alone does not prove artefact (UniProt Q9BSA9). Review morphology and detection controls before scoring (standard IHC practice). |
| Strong staining in adipocytes or colon glandular cells. | HPA reports TMEM175 as Not detected in those specific cell populations (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity, especially if the product also appears in a no-primary control (standard IHC practice). Avoid treating every cell in the tissue as an established negative. |
| Widespread, similarly intense staining across cells and tissue structures, with little compartment detail. | A uniform haze is difficult to reconcile with HPA's cell-specific staining levels and nuclear profile (HPA tissue IHC). It may reflect nonspecific antibody binding or detection background; compare the no-primary control and assess whether the stain follows intact cell boundaries (standard IHC practice). |
| No signal in caudate glial cells or testicular Sertoli cells. | Those are HPA's High reference populations (HPA tissue IHC). A blank result there does not establish biological absence. Check the tissue section, detection controls and assay conditions before interpreting a negative result (standard IHC practice); HPA's Approved rating still awaits external verification (HPA tissue IHC). |
💡Expected TMEM175 appearanceA reference-positive paraffin section shows High nuclear or nuclear-rim staining in caudate glial cells or testicular Sertoli cells (HPA tissue IHC); diffuse tissue-wide chromogen or similarly strong staining in HPA-negative cell populations warrants control review (HPA tissue IHC; standard IHC practice).
| Reference pattern and validation | HPA reports a nuclear and nuclear-membranous tissue IHC pattern and rates antibody HPA057160 Approved for IHC; its ICC rating is Uncertain (HPA tissue IHC; HPA antibodies). Approved here carries medium RNA–staining consistency and pending external verification, so the pattern is a reference for comparison rather than independent proof of molecular location (HPA tissue IHC). |
| Membrane topology and unknown epitope | TMEM175 has 12 transmembrane segments and alternating cytoplasmic and lumenal regions (UniProt Q9BSA9 topology). The supplied evidence does not map the antibody epitope, so topology cannot identify which side the antibody recognizes or predict a TMEM175-specific retrieval condition. |
| Expression breadth and cell-level controls | UniProt describes TMEM175 as widely expressed, and HPA assigns low tissue RNA specificity (UniProt Q9BSA9; HPA tissue IHC). Those broad descriptions do not override HPA's observed differences between named cell populations: select comparison regions by the reported cell type, not tissue name alone (HPA tissue IHC). |
| Antigen retrieval and assay conditions | Antigen retrieval can alter visibility in paraffin IHC, so document and optimize it with appropriate controls (standard IHC practice). Neither HPA tissue staining nor UniProt topology establishes a TMEM175-specific retrieval method or fixation sensitivity; do not infer either from a positive or negative tissue result. |
| Situation | Likely cause | Next action |
|---|---|---|
| The reference-positive caudate or testis section is blank. | HPA reports High staining in caudate glial cells and testicular Sertoli cells (HPA tissue IHC); a blank assay may reflect tissue selection or a failed detection step (standard IHC practice). | Confirm that the relevant cells are present, then check control reactions and review retrieval, primary-antibody dilution and detection conditions as general IHC variables (standard IHC practice). |
| Chromogen forms a diffuse haze over most of the section. | Uniform background obscures the cell and compartment pattern reported by HPA (HPA tissue IHC). Nonspecific binding or detection background are general IHC possibilities (standard IHC practice). | Compare a no-primary control, check blocking and washing, and adjust primary-antibody or detection conditions if controls indicate background (standard IHC practice). |
| Adipocytes or colon glandular cells stain strongly. | HPA reports those cell populations as Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity may account for an unexpected chromogenic result (standard IHC practice). | Check the no-primary control and inspect cell identity and compartment; repeat comparison against an HPA-positive cell population before assigning the signal to TMEM175 (HPA tissue IHC; standard IHC practice). |
| Staining appears only as cytoplasmic puncta, with no HPA-like nuclear pattern. | Puncta differ from HPA's tissue IHC profile but could fit the endosome/lysosome location in UniProt (HPA tissue IHC; UniProt Q9BSA9). The supplied evidence cannot resolve that discrepancy. | Document both observations, inspect assay controls and avoid calling puncta either confirmed TMEM175 or definite artefact from location alone (standard IHC practice). |
| Adjacent sections show inconsistent staining intensity. | HPA already notes medium consistency between tissue staining and RNA data (HPA tissue IHC). Section composition or variable assay conditions can also change apparent intensity (standard IHC practice). | Compare the same named cell population across sections and review section quality, retrieval, incubation and detection records before making a biological interpretation (standard IHC practice). |
| Q: Does a nuclear-rim IF/ICC image validate the paraffin IHC result? | HPA lists nucleoplasm and nuclear membrane as approved ICC-IF locations, but rates antibody HPA057160's ICC validation Uncertain (HPA subcellular; HPA antibodies). | A: Treat the IF/ICC image as a separate localisation observation. Compare the paraffin result with HPA's tissue IHC cell types and its Approved IHC pattern; neither readout resolves the conflict with UniProt's endosome/lysosome annotation (HPA tissue IHC; UniProt Q9BSA9). |
Experiment evidence
IHC & IF images showing TMEM175 staining
A11712-1 · IHCIHC analysis of TMEM175 using anti-TMEM175 antibody (A11712-1). TMEM175 was detected in a paraffin-embedded section of human prostate cancer
A11712-1 · IHCIHC analysis of TMEM175 using anti-TMEM175 antibody (A11712-1). TMEM175 was detected in a paraffin-embedded section of mouse testis tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000127419-TMEM175/tissue/cerebral+cortex#imid_18147059
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000127419-TMEM175/tissue/colon#imid_18147001
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000127419-TMEM175/tissue/liver#imid_18146994
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000127419-TMEM175/tissue/gallbladder#imid_18146970
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA057160; green = TMEM175, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA057160; green = TMEM175, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TMEM175 IHC & IF
🧪Run caudate first and assess staining in glial cells (High; HPA: caudate). Use adipose tissue adipocytes as the negative tissue (HPA: adipocytes Not detected). On the caudate slide, adjacent cells with absent or clearly weaker specific staining can serve as internal negative comparators if their identity and background are checked (standard IHC practice).
Positive control tissue: Caudate (Glial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TMEM175 in HEK293, MCF-7, U2OS, with annotated localisation: Nucleoplasm (approved), Nuclear membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype-matched control for the rabbit primary; and TMEM175 knockout tissue or cells where available as a biological negative (caption: rabbit primary; standard IHC practice). In caudate, quench endogenous peroxidase and inspect unstained sections for pigment that could complicate chromogenic scoring (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A11712-1 paraffin-section caption does not state the fixative (selected-SKU caption). That caption uses heat retrieval in EDTA at pH 8.0, but it does not establish whether retrieval is required under other conditions (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; for any IF follow-up, validate localization because HPA reports nucleoplasm and nuclear membrane in ICC-IF while UniProt places TMEM175 at endosome and lysosome membranes (HPA: subcellular; UniProt: subcellular location).
HPA tissue IHC evidence for TMEM175
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Testis | Sertoli cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | 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 → |
| Colon | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TMEM175 IHC Tips
Troubleshoot TMEM175 staining in paraffin sections using the catalog IHC conditions, cellular controls, and the reported compartment patterns.
Which retrieval conditions should I start with for TMEM175 paraffin-section IHC?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for TMEM175 paraffin-section IHC (datasheet A11712-1). The reported image used this retrieval, a 10% goat serum block, and 2 μg/ml primary antibody overnight at 4°C (datasheet A11712-1). If signal is weak, compare heating durations in the same buffer on adjacent sections while keeping detection and development constant (standard IHC practice). TMEM175 has 12 transmembrane segments and short connecting regions, so epitope accessibility may depend on retrieval conditions (UniProt Q9BSA9 topology). Check tissue integrity and no-primary controls before treating stronger DAB deposition as rescued specific staining (standard IHC practice).
Could fixation explain inconsistent TMEM175 staining between paraffin sections?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (datasheet A11712-1 caption). Record fixative, fixation duration, processing conditions, and section age before comparing specimens (standard IHC practice). Where histories differ, stain adjacent sections with the same EDTA pH 8.0 retrieval and 2 μg/ml primary incubation reported for the catalog image (datasheet A11712-1). Compare morphology, compartment pattern, and no-primary controls before interpreting weaker staining as lower TMEM175 abundance (standard IHC practice; UniProt Q9BSA9 localization). These data do not establish a TMEM175-specific effect of underfixation or overfixation (datasheet A11712-1 caption).
How should I assess nuclear staining when TMEM175 is a lysosomal membrane protein?
TMEM175 is assigned to endosome and lysosome membranes, making intracellular membrane-associated staining a biologically plausible IHC pattern (UniProt Q9BSA9 localization). The HPA tissue profile instead describes general nuclear and nuclear-membranous staining, and its subcellular record approves nucleoplasm and nuclear membrane locations (HPA tissue IHC; HPA subcellular). Treat a predominantly nuclear DAB pattern as an observation requiring validation, rather than assigning it automatically to lysosomes (UniProt Q9BSA9 localization; HPA subcellular). Compare adjacent sections with no-primary controls and an independent TMEM175 readout where available (standard IHC practice). Record nuclear and extranuclear staining separately so this compartment disagreement remains visible in the result (standard IHC practice).
Can this antibody distinguish TMEM175 isoforms or reveal which side of the membrane it binds?
TMEM175 has 2 annotated isoforms, but the supplied antibody caption does not map its epitope or establish isoform selectivity (UniProt Q9BSA9 isoforms; datasheet A11712-1 caption). Its 12 transmembrane segments separate cytoplasmic and lumenal regions, so accessibility cannot be inferred without the antibody's epitope sequence (UniProt Q9BSA9 topology). The annotated phosphothreonine at position 6 likewise does not establish whether this antibody detects a phosphorylation-dependent epitope (UniProt Q9BSA9 modified residues; datasheet A11712-1 caption). Test any claimed isoform specificity against mapped sequences and an appropriate independent control before interpreting compartment differences (standard IHC practice). Report the antibody identity and retrieval conditions with the staining result (datasheet A11712-1; standard IHC practice).
How can I use IF to check the cell type and compartment behind TMEM175 IHC staining?
Use a separate IF experiment to compare TMEM175 with a validated marker for the cell population being examined; HPA reports high staining in caudate glial cells (HPA tissue IHC). Choose a spectrally separated, preferably far-red TMEM175 channel when tissue autofluorescence interferes with shorter-wavelength signals (standard IF practice). Match permeabilisation to the antibody epitope: cytoplasmic regions and lumenal loops occupy different sides of TMEM175 membranes, while this antibody's epitope is unreported (UniProt Q9BSA9 topology; datasheet A11712-1 caption). Include single-label and no-primary controls to assess bleed-through and nonspecific fluorescence (standard IF practice). Compare the resulting compartment pattern with IHC without treating fluorescence colocalisation alone as proof of antibody specificity (standard IF practice).
What controls help separate TMEM175 DAB signal from nonspecific staining?
Begin with the reported 10% goat serum block and 2 μg/ml primary concentration, then adjust one condition at a time if background persists (datasheet A11712-1). Include a no-primary section and an endogenous-peroxidase block when using HRP and DAB (standard IHC practice). The selected image used a peroxidase-conjugated secondary for 30 minutes at 37°C and DAB development (datasheet A11712-1). Compare background with sections containing cells reported as not detected, such as adipocytes in adipose tissue, while recognizing that HPA calls its staining reliability Approved with medium RNA agreement (HPA tissue IHC). Assess diffuse deposits and tissue-edge staining separately from cell-associated signal (standard IHC practice).
How should I quantify TMEM175 IHC when staining varies by cell type and compartment? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring because TMEM175 has membrane localization in UniProt and nuclear-associated HPA staining (UniProt Q9BSA9 localization; HPA tissue IHC). For chromogenic sections, report the percentage of positive target cells and an intensity-based H-score from 0–300 using prespecified thresholds (standard IHC practice). If counting positive cells per area, report density per mm² of evaluable tissue and exclude folds or necrotic regions (standard IHC practice). Normalize counts to the number of eligible cells or evaluable area, using the same denominator across specimens (standard IHC practice). Keep nuclear and extranuclear scores separate and compare matched processing batches and no-primary controls (HPA subcellular; standard IHC practice).
When should I doubt that a TMEM175-positive IHC field reflects true target staining?
Question strong staining confined to compartments inconsistent with the proposed interpretation, since UniProt places TMEM175 on endosome and lysosome membranes while HPA reports nuclear-associated staining (UniProt Q9BSA9 localization; HPA tissue IHC). Check whether positive cells match the population under study; HPA reports high staining in caudate glial cells and no detection in adipocytes (HPA tissue IHC). Edge accentuation, necrotic areas, or staining reproduced in a no-primary section weaken a specific-signal interpretation (standard IHC practice). Residual endogenous peroxidase can also create DAB signal and should be assessed with appropriate controls (standard IHC practice). Resolve persistent compartment or cell-type discrepancies with an independent target readout before making biological claims (standard IHC practice).
Boster reagents
Best TMEM175 / Endosomal/lysosomal proton channel TMEM175 IHC Antibodies
The IHC-validated anti-TMEM175 antibody A11712-1 has paraffin-section images from human prostate cancer tissue and mouse and rat testis (catalog IHC captions).
Real IHC data
Anti-TMEM175 Antibody ®
Cat # A11712-1
A11712-1 is the only SKU that will render and lists IHC reactivity for human, mouse, and rat (catalog applications/reactivity). Its IHC captions document staining in paraffin sections of human prostate cancer tissue, mouse testis, and rat testis (catalog IHC captions).
Which to pick: Choose A11712-1 for paraffin-section IHC; its captions document EDTA pH 8.0 retrieval, 2 μg/ml primary antibody, and DAB detection, but do not report the fixative (catalog IHC captions). The same SKU covers human, mouse, and rat IHC (catalog applications/reactivity). No listed SKU has IF/ICC validation or an IF image, so this catalog does not support an IF/ICC recommendation (catalog applications and IF image records).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BSA9 (TM175_HUMAN, Endosomal/lysosomal proton channel TMEM175).
- Human Protein Atlas. TMEM175 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TMEM175 subcellular location (ICC-IF): Localized to the nucleoplasm and nuclear membrane..
- Human Protein Atlas. TMEM175 antibody validation summary (1 antibodies).
- Resveratrol attenuates inflammation and fibrosis in rheumatoid arthritis-associated interstitial lung disease via the AKT/TMEM175 pathway. Journal of translational medicine 2024 — PMC11095009.
- Common and Rare Variants in TMEM175 Gene Concur to the Pathogenesis of Parkinson's Disease in Italian Patients. Molecular neurobiology 2023 — PMC9984355.
- Deficiency of lysosomal TMEM175 in myeloid macrophages exerts anti-tumor immunity via inflammasome and cross-presentation pathway. Nature communications 2026 — PMC13018187.
- A growth-factor-activated lysosomal K(+) channel regulates Parkinson's pathology. Nature 2021 — PMC7979525.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:18691976 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TMEM175 (Endosomal/lysosomal proton channel TMEM175). Expected localisation Nuclear/rim in tissue (HPA tissue IHC); endolysosomal membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A11712-1). Validated IHC antibody: A11712-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A11712-1); 2. Fixation: Image fixative and duration unreported (datasheet A11712-1); 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 A11712-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A11712-1); 8. Primary antibody: Rabbit anti-TMEM175, 2-5 μg/ml (datasheet A11712-1); 9. Primary incubation: Overnight at 4 °C (datasheet A11712-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Nuclear tissue staining differs from the annotated membrane location (HPA tissue IHC; UniProt); Isoform / epitope: 2 isoforms ; map the epitope to lumenal or cytoplasmic regions (UniProt); Regulation: Staining-linked expression regulation is unreported (UniProt). Evidence: UniProt Q9BSA9, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and nuclear membrane., Datasheet A11712-1, PMC13018187. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TMEM175/immunohistochemistry-ihc-guide.