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UBTD1 / Ubiquitin domain-containing protein 1 · IHC design guide
Design Immunohistochemistry for UBTD1
Plan UBTD1 chromogenic IHC using mainly cytoplasmic tissue staining as a starting expectation (HPA tissue IHC). The catalog antibody's IHC dilution is 1:100–1:300 (datasheet); interpret staining with controls because HPA rates the tissue evidence uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBTD1 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 | Mainly cytoplasmic in several tissues (HPA tissue IHC) | |
| Staining pattern | Mainly cytoplasmic in adipocytes and squamous epithelia (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Appendix+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A16697-1) | |
| Caveat | Antibody staining and RNA expression show low consistency (HPA tissue IHC) | |
| Regulation | TP53 downstream target (UniProt) | |
| Isoform / epitope | No isoforms or processing annotated; epitope effects unknown (UniProt) |
Section 1
Recommended UBTD1 IHC & IF Protocols
Compare the catalog antibody's IHC-P protocol with the published UBTD1 colorectal tissue protocol (PMC11297151).
| Sample | Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A16697-1) |
| Fixation | Image fixative and duration unreported (datasheet A16697-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-UBTD1, 1:100 - 1:300 (datasheet A16697-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBTD1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Mainly cytoplasmic expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); the published protocol used microwave-heated sodium citrate at 95 °C (PMC11297151).
Section 2
What Is the Expected UBTD1 Staining Pattern?
In paraffin sections, expect mainly cytoplasmic UBTD1 staining, with High staining reported in adipocytes, cardiomyocytes, and selected epithelial and stromal cells (HPA tissue IHC). HPA rates the tissue IHC pattern Uncertain because staining has low consistency with RNA expression (HPA tissue IHC). UniProt does not annotate a subcellular location or a transmembrane segment, so it cannot independently confirm the observed compartment (UniProt Q9HAC8).
| Cytoplasmic staining in adipocytes from adipose tissue or breast, cardiomyocytes, or lung alveolar cells. | These are reported High cell populations and fit the mainly cytoplasmic tissue profile (HPA tissue IHC). Score the named cells rather than treating every cell in the section as an expected positive. The IHC evidence remains Uncertain (HPA tissue IHC). |
| Strong nuclear-only staining, or an exclusively sharp membrane outline, in otherwise expected-positive cells. | This does not match the mainly cytoplasmic tissue pattern (HPA tissue IHC). Review morphology, counterstain, and control sections before assigning UBTD1 positivity. HPA reports additional plasma-membrane localization in ICC-IF, so a membrane component alone cannot establish an artefact (HPA subcellular ICC-IF). |
| Prominent staining in appendix glandular cells or bone-marrow hematopoietic cells. | These cell populations were Not detected by HPA tissue IHC. Check whether the signal follows cell boundaries and persists after control checks; cross-reactivity or endogenous chromogen activity are possible general IHC explanations. Because HPA rates the IHC pattern Uncertain, this mismatch alone does not prove either cause (HPA tissue IHC). |
| Uniform pale color across stroma, empty spaces, and several unrelated cell types. | That distribution is harder to interpret as cell-specific UBTD1 staining than cytoplasmic signal in the named positive cells (HPA tissue IHC). In general chromogenic IHC practice, residual detection reagent, incomplete blocking, or excess primary antibody can produce diffuse background; compare a no-primary control. |
| No staining in adipocytes from adipose tissue despite an otherwise readable section. | Adipocytes are reported High in this tissue (HPA tissue IHC). A negative result merits a technical check, but does not by itself establish absent UBTD1: the tissue IHC reliability is Uncertain (HPA tissue IHC). Compare morphology and controls before changing the biological interpretation. |
💡Expected UBTD1 appearanceCall a section positive when named HPA High cell populations show predominantly cytoplasmic staining; uniform background or nuclear-only color is suspect, subject to the Uncertain IHC rating (HPA tissue IHC).
| Tissue and cell selection | HPA reports High staining in esophageal and oral squamous epithelium, nasopharyngeal respiratory epithelium, ovarian stroma, and several other listed cells, while selected glandular and hematopoietic cells are Not detected (HPA tissue IHC). Compare the specified cell type within each tissue. |
| Strength of the tissue evidence | The tissue pattern is rated Uncertain because antibody staining and RNA expression show low consistency; RNA tissue specificity is low (HPA tissue IHC). Treat a single positive or negative section as a provisional observation, particularly when the cell type differs from the HPA listing. |
| Topology and processing | UniProt lists no transmembrane segment, signal peptide, propeptide, or glycosylation site and describes one chain spanning residues 1–227 (UniProt Q9HAC8). These annotations provide no basis for predicting a secreted or shed staining pattern; they do not identify the antibody epitope. |
| Antibody-specific support | For HPA034825, HPA lists IHC as Uncertain and ICC as Approved (HPA antibodies). An ICC localization result helps frame a compartment question but does not upgrade validation of chromogenic paraffin-section staining. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive adipocytes show no chromogenic signal. | The section may have a detection or staining-workflow problem; HPA also rates tissue IHC Uncertain (HPA tissue IHC). | In general IHC practice, verify section integrity, retrieval and detection controls, and primary-antibody application. Recheck adipocytes in the section before concluding that the target is absent. |
| Signal fills the section rather than outlining identifiable cells. | Diffuse background can arise from nonspecific antibody binding or incomplete washing in general IHC practice. | Inspect a no-primary control and tissue morphology. Review blocking, wash steps, antibody concentration, and development time; retain only cell-resolved staining when comparing with HPA's cytoplasmic profile (HPA tissue IHC). |
| Color appears in an HPA Not detected cell population. | Cross-reactivity or endogenous detection activity is possible in general chromogenic IHC practice; HPA's negative listing is itself part of an Uncertain IHC profile (HPA tissue IHC). | Compare a no-primary control and check the detection system's endogenous-activity controls. Describe the observed cell type and compartment, then assess it alongside an HPA High cell population. |
| Most color is nuclear although the expected cells are present. | A nuclear-dominant pattern conflicts with HPA's mainly cytoplasmic tissue profile; UniProt supplies no location annotation to resolve it (HPA tissue IHC; UniProt Q9HAC8). | Recheck the counterstain and no-primary control, then compare the same cell type across sections. Record persistent nuclear staining as a discrepancy rather than scoring it as the expected pattern. |
| A weak signal is seen only in glandular cells. | Some glandular populations are Low, while appendix, epididymis, and parathyroid glandular cells are Not detected; the tissue and cell identity matter (HPA tissue IHC). | Identify the exact glandular population and compare it with the corresponding HPA entry. Use an expected-positive cell population as a staining reference before interpreting weak color. |
| Can ICC-IF images settle an ambiguous IHC compartment pattern? | HPA's ICC-IF shows mainly vesicles, with additional plasma membrane and cytosol, while tissue IHC is described as mainly cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). | Use the ICC-IF finding as context only. Judge the paraffin section by its cell-resolved tissue pattern and controls; HPA lists HPA034825 as ICC Approved but IHC Uncertain (HPA antibodies). |
Experiment evidence
IHC & IF images showing UBTD1 staining
A16697-1 · IHCImmunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue, using UBTD1 Antibody. The picture on the right is blocked w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000165886-UBTD1/tissue/cerebral+cortex#imid_8486444
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000165886-UBTD1/tissue/colon#imid_8486378
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000165886-UBTD1/tissue/liver#imid_8486363
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000165886-UBTD1/tissue/kidney#imid_8486372
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034825; green = UBTD1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034825; green = UBTD1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBTD1 IHC & IF
🧪Run adipose tissue first and require staining in adipocytes (HPA: High in adipocytes). Use appendix glandular cells as the negative tissue (HPA: Not detected); on the adipose slide, treat morphologically identifiable non-adipocytes as internal negative cells only if they remain unstained, since no specific internal negative cell type is reported.
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show UBTD1 in A-431, U-251MG, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus UBTD1 knockout material or a synthesized-peptide block as a specificity control (A16697-1 caption: peptide block). Quench endogenous peroxidase for chromogenic detection and assess staining around adipocytes rather than in empty lipid spaces (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; optimize retrieval empirically for paraffin sections. The A16697-1 paraffin-section caption does not report a fixative (A16697-1 caption: fixative unreported). Whether frozen sections or IF are easier is unreported; lipid spaces in adipose sections can complicate scoring (standard histology practice).
HPA tissue IHC evidence for UBTD1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBTD1 IHC Tips
Troubleshoot UBTD1 staining in paraffin sections using the catalog antibody’s tissue image, reported expression patterns, and matched controls.
How should I retrieve UBTD1 in paraffin sections when the signal is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min, for chromogenic IHC of paraffin sections (page retrieval setting). Keep section thickness, cooling time, antibody dilution, and detection conditions constant while comparing retrieval runs, because each can change apparent staining intensity (standard IHC practice). If staining remains weak, compare a second retrieval condition on adjacent sections as a fallback, recording tissue preservation and background alongside signal (standard IHC practice). Score cytoplasmic staining in the relevant cells rather than treating stronger staining anywhere in the section as improved retrieval (HPA: mainly cytoplasmic tissue expression; standard IHC practice).
Could fixation explain variable UBTD1 staining between paraffin blocks?
UBTD1-specific sensitivity to fixation is unknown here; the selected A16697-1 caption identifies paraffin-embedded colon carcinoma tissue but does not state its fixative (selected-SKU tissue-IHC caption). For a controlled comparison, process matched tissue pieces with the same fixative and compare defined fixation intervals, such as 6, 12, and 24 h, using identical embedding and staining conditions (standard IHC practice). Document cold ischemia, block age, section thickness, and retrieval run so these variables do not masquerade as fixation effects (standard IHC practice). Assess both cellular staining and morphology, then choose a condition that preserves interpretable tissue structure without raising nonspecific signal (standard IHC practice).
Should I score vesicular, membrane, or diffuse cytoplasmic UBTD1 staining?
For chromogenic tissue IHC, begin by scoring cytoplasmic staining within identified cells, since the reported tissue profile is mainly cytoplasmic (HPA: tissue IHC profile). Vesicles are the approved main location in cellular IF, with plasma membrane and cytosol as additional approved locations; that evidence helps frame hypotheses but does not establish a tissue-IHC scoring compartment (HPA: subcellular). UniProt supplies no subcellular annotation and lists no transmembrane segment, so a membrane rim alone needs separate validation before being called specific UBTD1 (UniProt Q9HAC8: subcellular annotation and topology). Record granular and diffuse patterns separately and compare each with omission and peptide-blocked controls (standard IHC practice; selected-SKU tissue-IHC caption).
How can I investigate an unexpected UBTD1 staining pattern when the epitope is unclear?
Map the antibody’s stated immunogen or epitope against the 227-aa UBTD1 sequence before interpreting a staining shift; the supplied record does not locate this antibody’s epitope (UniProt Q9HAC8: sequence length; supplied antibody evidence). The record lists a ubiquitin-like domain at residues 149–224, no listed isoforms, and no annotated glycosylation or modified residues, so none of those features alone explains a changed IHC pattern (UniProt Q9HAC8: domains, isoforms, modifications). The A16697-1 tissue caption includes a synthesized-peptide block image; compare that paired image with the unblocked stain when assessing antibody-dependent signal (selected-SKU tissue-IHC caption). Validate a disputed compartment with an independent antibody whose epitope is documented and with matched controls (standard IHC practice).
How should I translate the tissue IHC result into a multiplex UBTD1 IF experiment?
Use the chromogenic result to choose a cell population, then multiplex UBTD1 with a marker of that population; esophageal squamous epithelial cells are reported as high for UBTD1 (HPA: esophagus squamous epithelial cells). Select spectrally separated fluorophores, placing a weaker UBTD1 channel in the far red when tissue autofluorescence makes shorter wavelengths difficult to interpret, and include single-label controls (standard IF practice). Because the antibody epitope and its membrane-facing side are unspecified, compare staining with and without a mild permeabilisation step, such as 0.1% Triton X-100 for 5 min, before assigning intracellular access (supplied antibody evidence; standard IF practice). Compare vesicular, membrane, and cytosolic patterns with the reported cellular locations while keeping the IHC and IF readouts distinct (HPA: subcellular).
What controls help distinguish UBTD1 signal from chromogenic background?
Run a primary-antibody omission control and inspect adjacent tissue for diffuse chromogen, pigment, and staining at section edges (standard IHC practice). For peroxidase-based detection, a 3% hydrogen peroxide, 10 min block and controlled DAB development address endogenous enzyme signal and overdevelopment as general workflow steps (standard IHC practice). Compare the unblocked A16697-1 colon carcinoma image with its synthesized-peptide-blocked image when judging antibody-dependent staining; that caption does not specify fixation (selected-SKU tissue-IHC caption). If background persists, titrate the primary antibody and detection reagent on matched sections, then assess whether cytoplasmic signal remains in morphologically intact cells (HPA: mainly cytoplasmic tissue expression; standard IHC practice).
How should I quantify UBTD1 IHC across samples with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, using cytoplasmic signal as the initial tissue-IHC readout (HPA: mainly cytoplasmic tissue expression; standard IHC practice). Report the percentage of positive cells and intensity by category, or calculate an H-score from 0–300 using the percentages at intensity levels 0–3 (standard IHC practice). Normalise positive counts to the number of evaluable cells of the chosen type, or normalise stained area to that cell population’s measured area, rather than to whole-section area when composition varies (standard IHC practice). Keep exposure, threshold, retrieval, and scoring rules fixed, and report excluded folds, necrosis, and edges (standard IHC practice).
What makes a UBTD1-positive IHC result convincing rather than artefactual?
Prioritise staining in intact cells with a reproducible cytoplasmic pattern; tissue IHC reports mainly cytoplasmic expression, while cellular IF supports vesicles plus additional plasma membrane and cytosol localisation (HPA: tissue IHC and subcellular). Compare cell identity with the reported high-staining groups, such as esophageal squamous epithelial cells, and treat an isolated unexpected compartment or cell population as a finding requiring confirmation (HPA: esophagus squamous epithelial cells; standard IHC practice). Exclude section-edge signal, necrotic areas, pigment, and residual endogenous peroxidase activity through morphology and appropriate controls (standard IHC practice). Interpret concordance cautiously because HPA rates tissue-IHC reliability Uncertain owing to low consistency between antibody staining and RNA expression (HPA: tissue-IHC reliability).
Boster reagents
Best UBTD1 / Ubiquitin domain-containing protein 1 IHC Antibodies
A16697-1 has a paraffin-section IHC image from human colon carcinoma (catalog image caption); IF/ICC is listed without an IF image (catalog applications; catalog IF image alts). Human, mouse and rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-UBTD1 Antibody
Cat # A16697-1
A16697-1 has an IHC image of paraffin-embedded human colon carcinoma tissue with a peptide-blocked comparison (catalog image caption). The same SKU lists IHC, IF and ICC applications and human, mouse and rat reactivity; no IF image is provided (catalog applications; catalog reactivity; catalog IF image alts).
Which to pick: For tissue IHC, choose A16697-1 at the listed 1:100–1:300 range: its image documents paraffin-embedded human colon carcinoma, but the fixative is unreported (catalog IHC dilution; catalog image caption). For IF/ICC, A16697-1 lists both applications and is polyclonal, though no IF image is provided (catalog applications; catalog dilution record; catalog IF image alts). For mouse or rat samples, A16697-1 lists reactivity with both species, while its IHC image documents human tissue only (catalog reactivity; catalog image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9HAC8 (UBTD1_HUMAN, Ubiquitin domain-containing protein 1).
- Human Protein Atlas. UBTD1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. UBTD1 subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the plasma membrane and cytosol..
- Human Protein Atlas. UBTD1 antibody validation summary (1 antibodies).
- Ubiquitination-related Gene UBTD1 Mediates Poor Prognosis of Colorectal Cancer and Affects Colorectal Cancer Cell Proliferation and Ferroptosis. Recent patents on anti-cancer drug discovery 2026 — PMC13312376.
- CXCR4 mediates matrix stiffness-induced downregulation of UBTD1 driving hepatocellular carcinoma progression via YAP signaling pathway. Theranostics 2020 — PMC7255012.
- UBTD1: a prognostic and immune biomarker validated in thyroid cancer and pan-cancer analysis. Frontiers in oncology 2026 — PMC13388116.
- UBTD1 is a potential prognostic biomarker in colorectal cancer. Scientific reports 2024 — PMC11297151.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBTD1 (Ubiquitin domain-containing protein 1). Expected localisation Mainly cytoplasmic in several tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A16697-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Appendix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A16697-1); 2. Fixation: Image fixative and duration unreported (datasheet A16697-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-UBTD1, 1:100 - 1:300 (datasheet A16697-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A16697-1); Caveat: Antibody staining and RNA expression show low consistency (HPA tissue IHC); Isoform / epitope: No isoforms or processing annotated; epitope effects unknown (UniProt); Regulation: TP53 downstream target (UniProt). Evidence: UniProt Q9HAC8, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the plasma…, Datasheet A16697-1, PMC11297151. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBTD1/immunohistochemistry-ihc-guide.