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PSMD14 / Ubiquitin C-terminal hydrolase PSMD14 · IHC design guide
Design Immunohistochemistry for PSMD14
Plan PSMD14 paraffin IHC using the reported cytoplasmic and nuclear tissue pattern (HPA tissue IHC). Start with 2–5 μg/mL of catalog antibody A06584-1 (datasheet A06584-1), and interpret intensity cautiously because staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSMD14 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 nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Several tissues show cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06584-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Fallopian tube+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 | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Regulation is unspecified (UniProt) | |
| Isoform / epitope | No listed isoforms; one 1–310 chain (UniProt) |
Section 1
Recommended PSMD14 IHC & IF Protocols
The catalog antibody protocol (datasheet: A06584-1) is followed by four published PSMD14 IHC protocols (PMC10130002; PMC11165824; PMC13357097; PMC10688447).
| Sample | Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A06584-1) |
| Fixation | Image fixative and duration unreported (datasheet A06584-1); 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 A06584-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06584-1) |
| Primary antibody | Rabbit anti-PSMD14, 2-5 μg/ml (datasheet A06584-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06584-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06584-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSMD14-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A06584-1); published EDTA methods do not specify pH (PMC10130002; PMC13357097).
Section 2
What Is the Expected PSMD14 Staining Pattern?
In paraffin IHC, expect PSMD14 staining in the cytoplasm and nuclei of cells in several tissues (HPA tissue IHC). HPA reports medium staining in adipocytes, bone marrow hematopoietic cells, and cortical neurons, among other examples (HPA tissue IHC). Its tissue IHC reliability is Approved, with low consistency between antibody staining and RNA expression (HPA tissue IHC). PSMD14 has no annotated transmembrane segment (UniProt O00487 topology).
| Nuclear and cytoplasmic stain is visible in adipocytes or bone marrow hematopoietic cells. | This fits the reported compartment pattern and medium staining in those cell types (HPA tissue IHC). Judge signal in the named cells, since an overall tissue impression can hide which cells carry the stain (general IHC interpretation practice). |
| Stain is confined to membranes, extracellular material, or an unexpected structure. | That differs from HPA's cytoplasmic and nuclear tissue pattern and its supported nucleoplasmic ICC location (HPA tissue IHC; HPA subcellular). Check morphology and controls before assigning the unexpected compartment to PSMD14 (general IHC practice). |
| Strong stain appears in cells HPA lists as not detected, such as prostate glandular cells. | This conflicts with that specific HPA tissue observation (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible; a mismatch alone does not establish either cause (general IHC practice). Compare the suspect cells with controls and expected positive cells. |
| Color is spread across tissue, including spaces without intact cells. | Diffuse background obscures the reported cellular pattern (HPA tissue IHC). In chromogenic IHC, nonspecific binding, detection reagent activity, or residual chromogen can produce background (general IHC practice); the slide appearance alone cannot identify which occurred. |
| No stain is seen in an adequately sampled HPA medium-staining cell population. | An absent signal in, for example, adipocytes contrasts with HPA's medium observation (HPA tissue IHC). Check the assay and tissue integrity before calling PSMD14 absent; HPA also reports low staining in some cells and low staining–RNA consistency (HPA tissue IHC). |
💡Expected PSMD14 appearanceA convincing positive result is cellular nuclear and cytoplasmic stain in an HPA medium-staining population, such as adipocytes; diffuse color outside cells or dominant membrane-only stain is suspect (HPA tissue IHC; general IHC interpretation practice).
| Tissue and cell selection | HPA reports medium staining in several specified cell populations, low staining in others, and no detection in some; score the named cell type within its tissue (HPA tissue IHC). |
| Evidence strength | HPA marks tissue IHC Approved but notes low consistency with RNA expression; treat a single discrepant slide as a finding to verify, not a definitive expression map (HPA tissue IHC). |
| Antibody validation | HPA002114 has Approved IHC status; HPA003828 has Approved ICC status with no IHC status listed (HPA antibodies). Validation for one application does not establish performance in another (general assay practice). |
| Protein features | UniProt lists a full-length 1–310 chain, no signal peptide or propeptide, and no transmembrane segment (UniProt O00487). These features provide no basis for predicting an extracellular or membrane-only IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive cells are blank. | Assay failure, unsuitable tissue, or weak signal is possible (general IHC practice); HPA's medium category is an observation, not a guarantee for every section (HPA tissue IHC). | Confirm the named cells are present, review a positive control, and check the IHC-validated antibody's specified retrieval and detection steps (general IHC practice). |
| Color is widespread across the section. | Nonspecific binding or endogenous detection activity can obscure cellular staining (general IHC practice). | Inspect a negative control and the detection-only control; review blocking, washes, and chromogen development before scoring cells (general IHC practice). |
| Stain is strong in an HPA not-detected cell population. | The result disagrees with the listed observation; cross-reactivity or endogenous activity may be involved (HPA tissue IHC; general IHC practice). | Compare matched positive and negative controls, identify the stained cell type, and repeat the result before interpreting it as PSMD14 expression (general IHC practice). |
| Only membranes or extracellular material stain. | The compartment differs from HPA's nuclear and cytoplasmic tissue pattern (HPA tissue IHC); the appearance alone does not identify the source. | Check the counterstain and cellular boundaries, then compare with controls before accepting the localization (general IHC practice). |
| Does nucleoplasmic IF/ICC conflict with cytoplasmic and nuclear IHC? | HPA supports mainly nucleoplasmic localization in ICC-IF and describes cytoplasmic and nuclear expression in tissue IHC (HPA subcellular; HPA tissue IHC). | Interpret each observation in its own assay and cell context; use the tissue IHC pattern when judging a paraffin IHC slide (HPA tissue IHC; HPA subcellular). |
Experiment evidence
IHC & IF images showing PSMD14 staining
A06584-1 · IHCIHC analysis of POH1/PSMD14 using anti-POH1/PSMD14 antibody (A06584-1). POH1/PSMD14 was detected in a paraffin-embedded section of human gli
A06584-1 · IHCIHC analysis of POH1/PSMD14 using anti-POH1/PSMD14 antibody (A06584-1). POH1/PSMD14 was detected in a paraffin-embedded section of human lun
A06584-1 · IFIF analysis of POH1/PSMD14 using anti-POH1/PSMD14 antibody (A06584-1). POH1/PSMD14 was detected in an immunocytochemical section of U2OS cel
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115233-PSMD14/tissue/cerebral+cortex#imid_1051796
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115233-PSMD14/tissue/colon#imid_1051722
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115233-PSMD14/tissue/liver#imid_1051723
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000115233-PSMD14/tissue/kidney#imid_1051724
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002114; green = PSMD14, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002114; green = PSMD14, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSMD14 IHC & IF
🧪Run adipose tissue first and assess staining in adipocytes (HPA: Medium in adipocytes). Use fallopian tube glandular cells as the negative tissue (HPA: Not detected); HPA does not identify a negative cell type within the adipose section, so do not treat unstained neighboring cells as a validated internal negative.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Fallopian tube (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PSMD14 in A-431, U-251MG, U2OS, CACO-2, PC-3, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit isotype control; and a comparably processed PSMD14 knockout sample as a biological negative (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase for chromogenic detection and check adipose autofluorescence if evaluating IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A06584-1 caption does not state the fixative (caption: fixative not stated). The demonstrated paraffin-section procedure uses heat retrieval in EDTA at pH 8.0 before chromogenic detection; frozen sections and IF are not established as easier by the supplied evidence (caption: EDTA retrieval and DAB detection). Adipose lipid spaces leave thin cell rims to assess, so identify adipocyte nuclei and cell boundaries before scoring (standard tissue morphology).
HPA tissue IHC evidence for PSMD14
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PSMD14 IHC Tips
Troubleshoot PSMD14 staining in paraffin sections by checking retrieval, compartment-specific signal, background, and scoring against matched controls.
What retrieval should I try first when PSMD14 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06584-1). The selected image used 2 μg/ml primary antibody overnight at 4°C; keep those conditions fixed while comparing retrieval times on adjacent sections (datasheet A06584-1; standard IHC practice). If signal remains weak, test a shorter or longer heating interval, then consider a lower-pH buffer as a fallback while checking section integrity (standard IHC practice). Compare staining with a no-primary control and a reference containing cerebral-cortex neuronal cells, which showed medium staining in the tissue survey (standard IHC practice; HPA: Medium in cerebral-cortex neuronal cells).
Could fixation explain inconsistent PSMD14 staining between paraffin blocks?
The selected paraffin-section caption does not report a fixative, so PSMD14-specific fixation sensitivity is unknown (datasheet A06584-1). Record the fixative and fixation duration for each block, then compare matched sections using EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (datasheet A06584-1; standard IHC practice). If staining differs, assess morphology, section adherence, and nuclear accessibility before attributing the difference to antigen loss (standard IHC practice). Keep detection, DAB development time, and hematoxylin counterstaining consistent, and include no-primary controls to identify background associated with processing (standard IHC practice).
How should I assess nuclear versus cytoplasmic PSMD14 staining?
Expect predominantly nucleoplasmic signal in cell images, while tissue IHC has shown both nuclear and cytoplasmic expression across several tissues (HPA subcellular: Nucleoplasm supported; HPA tissue: Cytoplasmic and nuclear expression). Score the 2 compartments separately on well-preserved cells, using a hematoxylin counterstain to locate nuclei and a no-primary section to judge background (standard IHC practice). PSMD14 has no annotated transmembrane segment, but its UniProt record does not assign a subcellular location (UniProt O00487 topology and subcellular annotation). Investigate staining confined to tissue edges or damaged regions before interpreting a compartment shift, and compare sections developed for the same DAB time (standard IHC practice).
Could epitope accessibility explain variable PSMD14 staining?
The supplied record lists a 310-residue PSMD14 chain, an MPN domain at residues 31–166, and 0 annotated isoforms (UniProt O00487). It also lists phosphorylation at residues 150, 224, and 266, but gives no binding epitope for the catalog antibody (UniProt O00487 modified residues; datasheet A06584-1). Do not assign variable DAB intensity to an isoform or phosphorylation-dependent epitope without antibody-specific evidence (standard IHC interpretation). Compare adjacent sections with the documented EDTA pH 8.0 retrieval and consistent primary incubation before investigating an epitope-accessibility explanation (datasheet A06584-1; standard IHC practice).
How can I check PSMD14 localisation by multiplex IF?
Use IF as a separate localisation check: PSMD14 is mainly nucleoplasmic in the supplied cell images, and cerebral-cortex neuronal cells showed medium tissue staining (HPA subcellular: Nucleoplasm supported; HPA tissue: Medium in cerebral-cortex neuronal cells). Pair PSMD14 with a validated neuronal marker when examining that cell population, plus a nuclear counterstain, and assess overlap cell by cell (standard IF practice). Choose spectrally separated fluorophores, preferably a far-red PSMD14 channel if the specimen has strong shorter-wavelength autofluorescence, and include single-label controls (standard IF practice). Permeabilise sufficiently to access intracellular and nuclear epitopes while preserving morphology; PSMD14 has no annotated transmembrane segment, and the antibody epitope is unspecified (UniProt O00487 topology; datasheet A06584-1; standard IF practice).
What causes diffuse brown background in PSMD14 IHC?
First compare a no-primary section with the stained section: persistent DAB signal there suggests detection-system or tissue background rather than antibody-dependent PSMD14 staining (standard IHC practice). Quench endogenous peroxidase before HRP detection, control DAB development time, and check for pigment or precipitate under the microscope (standard IHC practice). The selected tissue image used 10% goat serum blocking, 2 μg/ml rabbit primary overnight at 4°C, and an HRP/DAB workflow (datasheet A06584-1). If background persists, check washing and compare a lower primary concentration on adjacent sections while retaining a tissue reference with expected staining (standard IHC practice; HPA: Medium in cerebral-cortex neuronal cells).
How should I quantify PSMD14 IHC across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analyse comparable, viable regions before scoring, since the tissue survey reports nuclear and cytoplasmic PSMD14 staining (HPA tissue: Cytoplasmic and nuclear expression; standard IHC practice). Report compartment-specific H-scores on a 0–300 scale, or the percentage of positive cells with a fixed threshold; use positive-cell density per mm² when area is the relevant denominator (standard IHC quantification). Normalise each score to the number of eligible cells or measured viable tissue area, and apply the same DAB exposure and image settings across cases (standard IHC practice). Record no-primary background and exclude folds, edges, and necrosis before comparing groups (standard IHC practice).
How do I distinguish genuine PSMD14 signal from staining artefact?
Look for reproducible cellular staining within intact tissue: PSMD14 is mainly nucleoplasmic in cell images, while the tissue survey also reports cytoplasmic staining (HPA subcellular: Nucleoplasm supported; HPA tissue: Cytoplasmic and nuclear expression). Compare the expected cell population with the no-primary control and a medium-staining reference such as cerebral-cortex neuronal cells (HPA: Medium in cerebral-cortex neuronal cells; standard IHC practice). Treat signal concentrated at edges, in necrosis, or in cell-free deposits as suspect, and check whether endogenous peroxidase contributes to DAB staining (standard IHC practice). Interpret weak or absent staining cautiously because the tissue survey flags low consistency between antibody staining and RNA expression (HPA tissue: Approved, low consistency).
Boster reagents
Best PSMD14 / Ubiquitin C-terminal hydrolase PSMD14 IHC Antibodies
A06584-1 has human paraffin-section IHC images and a U2OS-cell IF image (catalog image captions); both antibodies list human, mouse and rat reactivity (catalog reactivity).
Real IHC data
Anti-POH1/PSMD14 Antibody ®
Cat # A06584-1
A06584-1 has IHC images from human glioblastoma, lung adenocarcinoma, rectum adenocarcinoma and urothelial carcinoma paraffin sections, plus a U2OS-cell IF image (catalog image captions). M06584 lists IHC and IF/ICC applications for human, mouse and rat, but has no IHC or IF image in the payload (catalog applications/reactivity/image captions).
Which to pick: Choose A06584-1 for tissue IHC because its own captions document paraffin-section staining; the fixative is unreported (catalog IHC image captions). For IF/ICC, A06584-1 has a U2OS-cell IF image, while monoclonal M06584, clone 21P90, lists IF/ICC without an image (catalog IF image captions; catalog clone/applications). For cross-species work, both list human, mouse and rat reactivity, but the shown IHC results cover human tissue only (catalog reactivity; catalog IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00487 (PSDE_HUMAN, Ubiquitin C-terminal hydrolase PSMD14).
- Human Protein Atlas. PSMD14 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PSMD14 subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. PSMD14 antibody validation summary (2 antibodies).
- The prognostic role of PSMD14 in head and neck squamous cell carcinoma. Journal of cancer research and clinical oncology 2023 — PMC10130002.
- PSMD14 is a novel prognostic marker and therapeutic target in osteosarcoma. Diagnostic pathology 2024 — PMC11165824.
- Prognostic and therapeutic implications of PSMD14 in hepatocellular carcinoma: an integrative analysis of transcriptomic and single-cell profiles. Translational cancer research 2026 — PMC13357097.
- Regulation of Drp1 and enhancement of mitochondrial fission by the deubiquitinating enzyme PSMD14 facilitates the proliferation of bladder cancer cells. Oncology reports 2024 — PMC10688447.
- PubMed PMID:9374539 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSMD14 (Ubiquitin C-terminal hydrolase PSMD14). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06584-1). Validated IHC antibody: A06584-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Fallopian tube. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A06584-1); 2. Fixation: Image fixative and duration unreported (datasheet A06584-1); 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 A06584-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06584-1); 8. Primary antibody: Rabbit anti-PSMD14, 2-5 μg/ml (datasheet A06584-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06584-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No listed isoforms; one 1–310 chain (UniProt); Regulation: Regulation is unspecified (UniProt). Evidence: UniProt O00487, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet A06584-1, PMC10130002, PMC11165824. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMD14/immunohistochemistry-ihc-guide.