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DEPTOR / DEP domain-containing mTOR-interacting protein · IHC design guide
Design Immunohistochemistry for DEPTOR
Plan chromogenic DEPTOR IHC-P around variable cytoplasmic staining, with cardiomyocytes as a high-staining reference (HPA tissue IHC). This guide covers consistent fixation, the catalog antibody’s documented IHC-P workflow (datasheet A03811-2), and interpretation of tissue-dependent intensity (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DEPTOR 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 IHC (HPA tissue IHC); lysosomal membrane when mTOR-associated (UniProt) | |
| Staining pattern | Variable cytoplasmic staining; high in cardiomyocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03811-2) | |
| Positive control | Heart muscle+4 more · see all | |
| Negative control | Caudate+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 | Tissue staining varies; RNA concordance is medium (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; map the antibody epitope to each (UniProt) |
Section 1
Recommended DEPTOR IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet: A03811-2). Four published DEPTOR IHC methods provide paraffin-section conditions (PMC7471900; PMC5598754; PMC4657798; PMC4924707).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03811-2) |
| Fixation | Image fixative and duration unreported (datasheet A03811-2); 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 A03811-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03811-2) |
| Primary antibody | Rabbit anti-DEPTOR, 2-5 μg/ml (datasheet A03811-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03811-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03811-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DEPTOR-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression at variable levels in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A03811-2); the published methods give no alternative retrieval conditions.
Section 2
What Is the Expected DEPTOR Staining Pattern?
In paraffin-section IHC, expect variable cytoplasmic DEPTOR staining across tissues, with strong signal in cardiomyocytes, skeletal myocytes, placental syncytiotrophoblast cell bodies, and salivary glandular cells (HPA: tissue IHC). DEPTOR associates with lysosomal membranes through mTOR complexes and has no transmembrane segment (UniProt Q8TB45: localization and topology). HPA rates the tissue profile Enhanced, while noting medium staining–RNA consistency and pending external verification (HPA: tissue IHC).
| Clear cytoplasmic signal in the expected cells, strongest in heart muscle cardiomyocytes or skeletal muscle myocytes (HPA: High). | This fits the reported IHC distribution. Compare cells within the same section before scoring: HPA reports variable cytoplasmic expression across most tissues, so uniform intensity is not required (HPA: tissue IHC). |
| Predominantly nuclear or sharply surface-restricted chromogenic staining, with little cytoplasmic signal. | Treat this as discordant with the HPA tissue-IHC profile and check specificity and staining controls (HPA: cytoplasmic profile). UniProt places complex-associated DEPTOR at lysosomal membranes but reports no transmembrane segment; that annotation does not require visible lysosomal puncta in routine IHC (UniProt Q8TB45: localization and topology). |
| Strong staining in a cell population listed as Not detected, such as esophageal squamous epithelium (HPA: Not detected). | Consider cross-reactivity or endogenous detection activity; inspect a no-primary control and compare another IHC-validated antibody if available (standard IHC practice). A single positive field does not overturn the HPA cell-specific observation, especially given its pending external verification (HPA: tissue IHC). |
| Diffuse color over cells and surrounding section, without clear cellular boundaries. | This is background rather than a scorable DEPTOR pattern. Check blocking, washes, primary-antibody concentration, and the detection-only control (standard IHC practice). HPA describes cellular cytoplasmic staining, not a diffuse section-wide deposit (HPA: tissue IHC). |
| No signal in heart muscle cardiomyocytes despite interpretable tissue morphology (HPA: High). | The run needs review before a negative call. Confirm the IHC-validated antibody, tissue processing and retrieval used for that assay, reagent activity, and the positive-control slide (standard IHC practice). HPA's High category supports a useful comparison tissue; it does not guarantee every specimen will stain (HPA: tissue IHC). |
💡Expected DEPTOR appearanceCall a convincing IHC positive when cytoplasmic color is cell-associated and strong in an HPA High population such as cardiomyocytes; diffuse section-wide color or dominant nuclear staining is discordant and needs control review (HPA: tissue IHC; standard IHC practice).
| Cell type and tissue | Choose a reported High population for a positive comparison and a specified Not detected population for contrast; score the named cells, not an entire tissue as uniformly positive or negative (HPA: tissue IHC). |
| Localization and topology | The IHC readout is cytoplasmic (HPA: tissue IHC). Lysosomal-membrane association depends on mTOR-complex association, and DEPTOR lacks a transmembrane segment; neither fact predicts a crisp membrane outline in chromogenic sections (UniProt Q8TB45: localization and topology). |
| Antibody evidence | HPA lists Enhanced IHC evidence for HPA023938, HPA023945, and CAB020841, and Supported IHC evidence for HPA024011 (HPA: antibodies). The tissue profile still has medium staining–RNA consistency and awaits external verification (HPA: tissue IHC). |
| Isoforms and modified residues | UniProt lists two isoforms and multiple modified residues, including phosphorylation sites (UniProt Q8TB45: isoforms and modified residues). Their effects on a particular antibody's IHC signal cannot be assigned without its epitope and validation data; no target-specific fixation sensitivity is supplied. |
| IF/ICC Q&A: should it match the IHC compartment exactly? | No direct match is established. HPA reports mainly mitochondrial ICC-IF signal, with additional nucleoplasm and nuclear-body signal, while its tissue IHC profile is cytoplasmic (HPA: subcellular ICC-IF; HPA: tissue IHC). Interpret each assay with its own validation and controls (standard IHC/IF practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive comparison tissue is blank. | A failed staining run, inactive detection reagents, or an unsuitable assay setup is possible (standard IHC practice). | Check the IHC-validated antibody's instructions, retrieval and detection steps, and a known working control before interpreting DEPTOR absence (standard IHC practice; HPA: High in cardiomyocytes). |
| All cells show similar brown color. | Excess background or endogenous detection activity may obscure the expected cell-specific pattern (standard IHC practice; HPA: variable cytoplasmic expression). | Run no-primary and detection-only controls; review blocking, washes, and primary concentration for the chosen detection system (standard IHC practice). |
| A reported Not detected cell population stains strongly. | Cross-reactivity, endogenous activity, or a specimen-specific difference may explain discordance (standard IHC practice; HPA: Not detected categories). | Verify cell identity and controls, then compare staining with an independent IHC-validated antibody where feasible; record the discrepancy rather than calling it established DEPTOR expression (HPA: antibodies; standard IHC practice). |
| Signal is nuclear with little cytoplasmic staining. | This differs from the HPA tissue-IHC profile; ICC-IF nuclear annotations alone do not validate that chromogenic IHC result (HPA: tissue IHC; HPA: subcellular ICC-IF). | Review controls, tissue morphology, and antibody-specific IHC evidence; score the nuclear pattern separately until its specificity is established (standard IHC practice). |
| Staining appears weak in a low-expression comparison tissue. | HPA already reports Low staining in populations such as cerebral-cortex endothelial cells and duodenal glandular cells (HPA: tissue IHC). | Compare with a reported High population in the same validated workflow before changing retrieval or dilution; document cell type and intensity separately (HPA: tissue IHC; standard IHC practice). |
| IHC looks diffuse, but an ICC-IF image looks punctate. | The reported readouts differ: cytoplasmic tissue IHC versus mainly mitochondrial ICC-IF; morphology and assay context also affect what can be resolved (HPA: tissue IHC; HPA: subcellular ICC-IF; standard microscopy practice). | Judge the paraffin-section result against tissue-IHC controls and the correct cells. Use the ICC-IF annotation as context, not as a required punctate IHC scoring rule (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing DEPTOR staining
A03811-2 · IHCIHC analysis of DEPDC6/DEPTOR using anti-DEPDC6/DEPTOR antibody (A03811-2). DEPDC6/DEPTOR was detected in a paraffin-embedded section of hum
A03811-2 · IHCIHC analysis of DEPDC6/DEPTOR using anti-DEPDC6/DEPTOR antibody (A03811-2). DEPDC6/DEPTOR was detected in a paraffin-embedded section of hum
A03811 · IHCImmunohistochemistry of DEPTOR in mouse liver tissue with DEPTOR antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000155792-DEPTOR/tissue/cerebral+cortex#imid_6264728
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000155792-DEPTOR/tissue/colon#imid_6273438
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000155792-DEPTOR/tissue/liver#imid_6264367
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000155792-DEPTOR/tissue/kidney#imid_5433940
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB020841; green = DEPTOR, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB020841; green = DEPTOR, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DEPTOR IHC & IF
🧪Run salivary gland first: glandular cells should stain (HPA: High in salivary gland glandular cells). Use esophagus squamous epithelial cells as the negative tissue (HPA: Not detected); on the positive slide, compare glandular cells with morphologically distinct stromal cells that lack specific DAB staining, treating those cells as candidate internal negatives because their DEPTOR status is not specified (HPA: salivary gland glandular-cell row).
Positive control tissue: Heart muscle (Cardiomyocytes, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DEPTOR in A-431, U-251MG, U2OS, RPTEC/TERT1, MCF-7, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species-matched isotype controls, plus DEPTOR-knockout material or a validated immunizing-peptide block as a biological specificity control (standard IHC practice). Quench endogenous peroxidase and inspect the salivary gland section for background DAB signal before scoring (standard chromogenic IHC practice).
⚠️Feasibility: The selected paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; its fixative and any target-specific fixation window or effect are unreported (A03811-2 tissue-IHC caption). Frozen sections and IF cannot be judged easier from the supplied tissue-IHC evidence; the HPA IF images are from cell lines (HPA: subcellular ICC-IF images). Assess glandular luminal material for nonspecific chromogen accumulation when scoring the salivary gland (standard IHC practice).
HPA tissue IHC evidence for DEPTOR
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Placenta | Syncytiotrophoblasts - cell body | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DEPTOR IHC Tips
Use the catalog antibody’s paraffin-section IHC conditions as a starting point, then judge DEPTOR staining by cell type, compartment and appropriate controls.
Which retrieval conditions should I try first for weak DEPTOR staining in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03811-2). The selected tissue-IHC image used that retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A03811-2). If staining remains weak, optimize heating duration and cooling consistently across control and test sections, while checking that the primary antibody and DAB detection steps worked (standard IHC practice). Compare the resulting cytoplasmic signal with expected cell-type patterns, such as high staining in cardiomyocytes or skeletal myocytes, rather than judging intensity alone (HPA tissue IHC).
How should I troubleshoot fixation when DEPTOR staining is weak or uneven?
The selected paraffin-section caption does not report a fixative, so DEPTOR-specific sensitivity to fixation is unknown (datasheet A03811-2). Record the fixative, fixation duration, tissue thickness and processing history for each specimen before attributing a weak result to antigen loss (standard IHC practice). Run similarly processed control and test sections together using EDTA at pH 8.0 and the same staining conditions to separate processing variation from assay variation (datasheet A03811-2; standard IHC practice). If staining tracks fixation batches, adjust processing systematically and assess morphology alongside signal; HPA expression patterns do not establish a fixation mechanism (standard IHC practice; HPA tissue IHC).
Where should DEPTOR staining appear, and how should I assess unexpected nuclear signal?
Expect variable cytoplasmic staining in tissue sections, with cell-type differences across organs, when assessing chromogenic IHC (HPA tissue IHC). DEPTOR can localize to the lysosomal membrane when associated with mTOR complexes, despite having no transmembrane segment (UniProt Q8TB45 subcellular location and topology). Separate HPA cell-imaging data place it mainly in mitochondria, with additional nucleoplasm and nuclear-body signal, so nuclear staining alone should not establish tissue-IHC specificity (HPA subcellular; standard IHC practice). Review nuclear and cytoplasmic signal against morphology, a matched negative control and expected positive cells before assigning a compartmental pattern (standard IHC practice; HPA tissue IHC).
Could DEPTOR isoforms or modifications explain different staining patterns?
DEPTOR has 2 annotated isoforms and DEP and PDZ domains, but the supplied caption does not identify the catalog antibody’s epitope (UniProt Q8TB45 isoforms and domains; datasheet A03811-2). Consequently, a difference between tissue samples cannot be assigned to isoform selectivity without an epitope map or independent validation (standard IHC practice). DEPTOR also has annotated phosphorylation sites, including serine 235, threonine 241 and serine 244, but their effect on this antibody’s staining is unknown (UniProt Q8TB45 modified residues; datasheet A03811-2). If patterns disagree, compare staining under identical retrieval and detection conditions, then investigate epitope coverage before proposing a biological mechanism (standard IHC practice).
How can I investigate DEPTOR localisation by IF alongside this IHC assay?
Treat IF/ICC as a separate assay: the selected catalog evidence describes chromogenic staining of a paraffin section, not an IF validation or fixation condition (datasheet A03811-2). For multiplex imaging, pair DEPTOR with a marker that identifies the expected cell type and use separated fluorophore channels chosen after checking tissue autofluorescence (HPA tissue IHC; standard IF practice). Because DEPTOR has no transmembrane segment and can associate with the lysosomal membrane, establish antibody access to the relevant intracellular epitope with controlled permeabilisation rather than assuming an extracellular-facing target (UniProt Q8TB45 topology and subcellular location; standard IF practice). Confirm any punctate overlap with single-stain and secondary-only controls before interpreting colocalisation (standard IF practice).
What should I check when DAB background obscures DEPTOR staining?
First compare the primary-antibody section with a no-primary control to locate signal arising from secondary reagent, detection chemistry or tissue rather than specific DEPTOR binding (standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody, a peroxidase-conjugated secondary and DAB development (datasheet A03811-2). If diffuse staining persists, optimize blocking, primary concentration, washing and DAB development while keeping retrieval at EDTA pH 8.0 for the initial comparison (datasheet A03811-2; standard IHC practice). Include a general endogenous-peroxidase blocking step and inspect pigment or precipitate before scoring cytoplasmic staining (standard IHC practice; HPA tissue IHC).
How should I score DEPTOR IHC across samples with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring area before analysis because DEPTOR staining varies by tissue and cell type (HPA tissue IHC; standard IHC practice). For cell-rich regions, report the percentage of positive target cells and an H-score using intensity categories 0–3; for spatial counts, report positive cells per mm² of viable tissue (standard IHC practice). Normalize comparisons to the number of eligible cells or viable tissue area, using the same segmentation rules, retrieval and DAB exposure across sections (standard IHC practice). Record compartment and staining threshold separately, and avoid treating stromal or necrotic areas as equivalent to the target-cell population (standard IHC practice).
How do I distinguish convincing DEPTOR staining from section artefacts?
A convincing result follows cell morphology and a reproducible compartmental pattern, with cytoplasmic tissue staining interpreted against known cell-type differences (HPA tissue IHC; standard IHC practice). High signal in cardiomyocytes or skeletal myocytes can support assay performance, whereas staining in a cell population reported as undetected warrants closer control review (HPA tissue IHC). Examine unexpected nuclear-only signal, section-edge enhancement, necrotic regions and granular pigment before calling DEPTOR positive; cell-imaging nuclear localisation alone does not validate a tissue-IHC pattern (HPA subcellular; standard IHC practice). Compare no-primary and endogenous-peroxidase controls with the stained section to identify DAB signal unrelated to antibody binding (standard IHC practice).
Boster reagents
Best DEPTOR / DEP domain-containing mTOR-interacting protein IHC Antibodies
Two anti-DEPTOR antibodies have real IHC images: human paraffin sections for A03811-2 and mouse liver for A03811 (catalog image captions). Neither has IF data (catalog application lists and images).
Real IHC data
Anti-DEPDC6/DEPTOR Antibody ®
Cat # A03811-2
Real IHC data
Anti-DEPTOR Antibody
Cat # A03811
A03811-2 shows IHC in human lung and stomach cancer paraffin sections (catalog image captions). A03811 shows IHC in mouse liver (catalog image caption).
Which to pick: Choose A03811-2 for human paraffin-section IHC; its caption reports EDTA retrieval and 2 μg/ml primary antibody, but does not report the fixative (A03811-2 image caption). Choose A03811 for mouse liver IHC at 5 μg/mL (A03811 image caption). Both list human, mouse and rat reactivity, but their pictured IHC results cover only the samples above; neither lists IF/ICC or provides an IF image, so neither is established here for IF/ICC (catalog reactivity, application lists and images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TB45 (DPTOR_HUMAN, DEP domain-containing mTOR-interacting protein).
- Human Protein Atlas. DEPTOR tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DEPTOR subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm and nuclear bodies..
- Human Protein Atlas. DEPTOR antibody validation summary (4 antibodies).
- Metformin attenuates renal interstitial fibrosis through upregulation of Deptor in unilateral ureteral obstruction in rats. Experimental and therapeutic medicine 2020 — PMC7471900.
- Downregulation of DEPTOR inhibits the proliferation, migration, and survival of osteosarcoma through PI3K/Akt/mTOR pathway. OncoTargets and therapy 2017 — PMC5598754.
- High DEPTOR expression correlates with poor prognosis in patients with esophageal squamous cell carcinoma. OncoTargets and therapy 2015 — PMC4657798.
- DEPTOR suppresses the progression of esophageal squamous cell carcinoma and predicts poor prognosis. Oncotarget 2016 — PMC4924707.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DEPTOR (DEP domain-containing mTOR-interacting protein). Expected localisation Cytoplasmic IHC (HPA tissue IHC); lysosomal membrane when mTOR-associated (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03811-2). Validated IHC antibody: A03811-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03811-2); 2. Fixation: Image fixative and duration unreported (datasheet A03811-2); 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 A03811-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03811-2); 8. Primary antibody: Rabbit anti-DEPTOR, 2-5 μg/ml (datasheet A03811-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03811-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Tissue staining varies; RNA concordance is medium (HPA tissue IHC); Isoform / epitope: 2 isoforms; map the antibody epitope to each (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q8TB45, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A03811-2, PMC7471900, PMC5598754. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DEPTOR/immunohistochemistry-ihc-guide.