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PSMD1 / 26S proteasome non-ATPase regulatory subunit 1 · IHC design guide
Design Immunohistochemistry for PSMD1
Plan chromogenic PSMD1 IHC in paraffin sections with the catalog antibody at 2–5 μg/ml (datasheet A06293-1). Assess cytoplasmic and nuclear staining, using kidney tubule cells as a high staining reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSMD1 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 | Cytoplasm and nucleus in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining across most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06293-1) | |
| Positive control | Breast+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; no transmembrane segment; epitope coverage unknown (UniProt) |
Section 1
Recommended PSMD1 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A06293-1). Two published PSMD1 IHC protocols provide additional settings for spinal cord sections and tumor tissue microarrays (PMC13538453; PMC10725586).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A06293-1) |
| Fixation | Image fixative and duration unreported (datasheet A06293-1); 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 A06293-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06293-1) |
| Primary antibody | Rabbit anti-PSMD1, 2-5 μg/ml (datasheet A06293-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06293-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06293-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSMD1-positive staining in myoepithelial cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A06293-1). Citrate pH 6.0 is a published alternative for spinal cord sections (PMC13538453).
Section 2
What Is the Expected PSMD1 Staining Pattern?
Expect PSMD1 staining in the cytoplasm and nuclei of many cell types, with strong staining in selected cells such as kidney tubule cells and cerebellar Purkinje cells (HPA: tissue IHC). PSMD1 has no transmembrane segment (UniProt Q99460: topology). HPA rates tissue staining reliability as Enhanced, while reporting medium agreement with RNA data and pending external verification (HPA: tissue IHC).
| Cytoplasmic and nuclear staining appears across much of a section, with strong kidney tubule staining. | This fits the broad tissue pattern and a documented high signal in tubule cells (HPA: tissue IHC). Judge the named cells and compartments together; staining throughout a section alone does not identify PSMD1. |
| Signal is confined to membranes or appears only outside cells. | That distribution conflicts with the reported cytoplasmic and nuclear tissue pattern (HPA: tissue IHC) and PSMD1's lack of a transmembrane segment (UniProt Q99460: topology). Review morphology, background and detection before calling it specific staining (general IHC practice). |
| Lung alveolar cells stain strongly while expected positive cells are weak. | Alveolar cells are reported as low, whereas kidney tubule cells and several other listed cell types are high (HPA: tissue IHC). Consider cross-reactivity or endogenous detection activity, but do not infer either cause from this contrast alone (general IHC practice). |
| Color spreads over tissue and empty areas, obscuring cell boundaries. | Diffuse background prevents a dependable compartment call (general IHC practice). Compare a control lacking primary antibody, then assess blocking, detection and washing; background cannot be assigned to PSMD1 from appearance alone (general IHC practice). |
| A kidney section shows no tubule signal. | Tubule cells are a documented high staining population (HPA: tissue IHC). Check tissue quality, antibody application and detection controls before treating the absence as biological; a single negative section cannot establish absent PSMD1 (general IHC practice). |
💡Expected PSMD1 appearanceA convincing positive shows cytoplasmic and nuclear staining with strong signal in a documented high population such as kidney tubule cells (HPA: tissue IHC); isolated membrane color or diffuse cell-free color calls for artefact checks (UniProt Q99460: topology; general IHC practice).
| Tissue and cell choice | Kidney tubule cells, Purkinje cells, cardiomyocytes and breast myoepithelial cells are reported high; lung alveolar cells are low (HPA: tissue IHC). Choose the positive population before scoring, and treat low staining as a comparison rather than proof of a true negative. |
| Compartment and complex | Tissue IHC reports cytoplasmic and nuclear expression in most tissues (HPA: tissue IHC). PSMD1 is a 19S proteasome regulatory particle component with no transmembrane segment (UniProt Q99460: subunit, topology); complex membership alone does not define a narrower staining location. |
| Validation strength | HPA036736 and HPA036737 have Enhanced IHC status; CAB021092 has Approved IHC status (HPA: antibody validation). HPA also reports medium antibody and RNA agreement with external verification pending (HPA: tissue IHC). Interpret an unexpected pattern with those limits in view. |
| Isoforms and processing | Two isoforms are listed, and the annotated chain spans residues 1–953 with no signal peptide or propeptide (UniProt Q99460: isoforms, processing). These facts do not establish which isoform an antibody detects or predict an IHC staining difference; check the antibody's documented target region if available. |
| IF/ICC Q: Should the same pattern be expected? | HPA places the main IF/ICC signal in the nucleoplasm and additionally at actin filaments (HPA: subcellular ICC-IF). That is an IF/ICC observation, while tissue IHC reports cytoplasmic and nuclear staining (HPA: tissue IHC); assess each application against its own evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in kidney tubule cells. | An assay step may have failed; tubule cells are reported high (HPA: tissue IHC). | Inspect a known-positive section and detection controls, then review antibody dilution and antigen retrieval against the chosen antibody's IHC-P instructions (general IHC practice). |
| Only membrane outlines stain. | This does not match the tissue compartment pattern (HPA: tissue IHC) or lack of a transmembrane segment (UniProt Q99460: topology). | Compare a control lacking primary antibody and reassess the chromogen deposit against cell boundaries before assigning localisation (general IHC practice). |
| Low-reporting cells dominate the section. | Strong alveolar staining is unexpected relative to the reported low level in those cells (HPA: tissue IHC); specificity or detection background needs checking. | Compare a documented high population on a control section and include a control lacking primary antibody; investigate any signal that persists without primary antibody (general IHC practice). |
| Diffuse color obscures nuclei and cytoplasm. | Background or excessive detection signal may prevent compartment scoring (general IHC practice). | Review blocking, washing, antibody dilution and chromogen development; score PSMD1 only where cellular boundaries and compartments remain interpretable (general IHC practice). |
| Only nuclear signal appears in tissue IHC. | HPA reports both nuclear and cytoplasmic tissue staining, although intensity can differ among cells (HPA: tissue IHC). | Check the cytoplasmic compartment in documented high cells and compare a control section before concluding the pattern is restricted to nuclei (general IHC practice). |
| Actin-like IF/ICC signal is expected in the IHC section. | Actin filament localisation is an additional ICC-IF finding; the tissue IHC summary describes cytoplasmic and nuclear expression (HPA: subcellular ICC-IF; HPA: tissue IHC). | Score the chromogenic section using tissue IHC compartments and cell populations; consult the separate IF/ICC guide for that application's localisation (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing PSMD1 staining
A06293-1 · IHCIHC analysis of PSMD1 using anti-PSMD1 antibody (A06293-1). PSMD1 was detected in a paraffin-embedded section of human placenta tissue. Heat
A06293-1 · IHCIHC analysis of PSMD1 using anti-PSMD1 antibody (A06293-1). PSMD1 was detected in a paraffin-embedded section of human bladder cancer tissue
A06293-1 · IFIF analysis of PSMD1 using anti-PSMD1 antibody (A06293-1) and anti-Beta Tubulin antibody (M01857-3). PSMD1 was detected in an immunocytochem
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000173692-PSMD1/tissue/cerebral+cortex#imid_15360406
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000173692-PSMD1/tissue/colon#imid_15360340
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000173692-PSMD1/tissue/liver#imid_15360333
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000173692-PSMD1/tissue/kidney#imid_15360326
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036736; green = PSMD1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036736; green = PSMD1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSMD1 IHC & IF
🧪Run breast first and score staining in myoepithelial cells (HPA: High in breast myoepithelial cells). HPA lists no negative tissue because PSMD1 is detected in all 45 scored tissues, so use no-primary and isotype controls; no cell type on the positive slide is established as an internal biological negative (HPA: no negative tissue listed).
Positive control tissue: Breast (Myoepithelial cells, HPA High)
Negative control tissue: None in HPA: PSMD1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PSMD1 in A-431, MCF-7, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a secondary-only slide and a matched nonimmune rabbit IgG isotype control (caption: rabbit anti-PSMD1 antibody), plus PSMD1-knockout material as a biological negative where available. Quench endogenous peroxidase and check for residual background on the secondary-only breast section before scoring HRP/DAB staining (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A06293-1 tissue-IHC caption does not state a fixative (caption: fixative not stated). The paraffin-section example uses heat-mediated EDTA retrieval at pH 8.0, but it does not establish that retrieval is required or that frozen sections or IF are easier (caption: EDTA retrieval in a paraffin section). In breast tissue, the thin myoepithelial layer requires careful assignment of staining to cells bordering the ducts (HPA: High in breast myoepithelial cells; standard histology).
HPA tissue IHC evidence for PSMD1
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 |
|---|---|---|---|---|
| Breast | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PSMD1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PSMD1 IHC Tips
Troubleshoot PSMD1 staining in paraffin sections by checking retrieval, intracellular localisation, controls and cell-specific scoring.
What retrieval should I try first when PSMD1 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A06293-1). The selected paraffin-section image used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C (caption A06293-1). If staining remains weak, vary heating duration on adjacent sections while keeping the antibody concentration and detection conditions constant (standard IHC practice). Include a known positive section in each run so a retrieval failure is distinguishable from low expression in the specimen (standard IHC practice). Assess tissue preservation alongside signal, since excessive heating can damage morphology and make compartment scoring unreliable (standard IHC practice).
Can I infer an optimal fixative for PSMD1 from the available tissue staining?
No target-specific fixation sensitivity is established here: the selected PSMD1 image identifies a paraffin-embedded section but does not state its fixative (caption A06293-1). Record the actual fixative and fixation interval for each specimen, then compare sections processed under matched retrieval and detection conditions (standard IHC practice). If nuclear or cytoplasmic staining changes across processing batches, assess morphology and a positive control before attributing the difference to PSMD1 abundance (standard IHC practice). Keep antibody concentration at the documented 2 μg/ml during that comparison (caption A06293-1). The available tissue pattern and protein annotations do not establish how PSMD1 responds to fixation (HPA tissue IHC; UniProt Q99460).
Should PSMD1 appear in nuclei, cytoplasm, or both on chromogenic IHC?
Score nuclear and cytoplasmic staining separately: tissue IHC reports both compartments in most tissues (HPA tissue IHC). Cell imaging places PSMD1 mainly in the nucleoplasm, with additional actin-filament localisation, so a purely membranous outline warrants scrutiny (HPA subcellular). PSMD1 has no annotated transmembrane segment, which gives no basis for treating membrane-only staining as the expected pattern (UniProt Q99460 topology). Compare the compartments within morphologically intact cells on the same section, using a counterstain to identify nuclei (standard IHC practice). If one compartment appears only at section edges or in damaged areas, repeat staining with matched retrieval before assigning biological meaning (standard IHC practice).
Could PSMD1 isoforms or modifications explain a missing IHC signal?
PSMD1 has 2 annotated isoforms, but this payload does not map the catalog antibody’s epitope to either one (UniProt Q99460; caption A06293-1). It also records modifications including phosphorylation at residues 273 and 290 and acetylation at residue 310; their effects on this antibody’s staining are unknown (UniProt Q99460). Before invoking isoform or modification dependence, verify retrieval, primary incubation and positive-control staining on the same run (standard IHC practice; caption A06293-1). If an epitope sequence becomes available, compare it with both isoforms and the annotated modification sites (UniProt Q99460). Report any resulting isoform claim as conditional until the antibody’s reactivity is established.
How can IF help check an ambiguous PSMD1 IHC pattern?
Use IF as a separate cross-check of compartment and cell identity, since tissue IHC reports nuclear and cytoplasmic PSMD1 while cell imaging reports mainly nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). Multiplex with a validated marker for the expected cell population, such as a Purkinje-cell marker when examining cerebellum, and compare signal within those cells (HPA tissue IHC; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence, favoring a spectrally separated red or far-red channel when background warrants it (standard IF practice). Because PSMD1 lacks a transmembrane segment and its antibody epitope is unmapped, optimise permeabilisation for intracellular access without assuming a membrane-facing epitope (UniProt Q99460 topology; standard IF practice).
How do I distinguish diffuse PSMD1 staining from chromogenic background?
Use a no-primary control to reveal secondary-reagent or detection background, and inspect a positive section in the same run (standard IHC practice). Block endogenous peroxidase before HRP and DAB development, particularly when unexplained pigment appears outside intact cells (standard IHC practice). The documented image used 10% goat serum blocking, an HRP-linked secondary incubation for 30 minutes at 37°C, and DAB development (caption A06293-1). Assess whether signal follows intact nuclear or cytoplasmic compartments, as reported for PSMD1 in tissue, rather than tissue folds or cut edges (HPA tissue IHC; standard IHC practice). Reduce nonspecific staining by checking wash adequacy before changing the documented primary concentration (standard IHC practice; caption A06293-1).
What should I quantify when PSMD1 stains both nuclei and cytoplasm? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear and cytoplasmic compartments separately because both occur in the reported tissue IHC pattern (HPA tissue IHC). For each compartment, record the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories for a 0–300 summary (standard IHC practice). If spatial distribution matters, report positive-cell density per mm² of viable tissue as well (standard IHC practice). Normalise counts to the number of eligible cells or measured viable area, and apply the same threshold to all comparison sections (standard IHC practice). Keep retrieval, development and imaging settings consistent across batches before comparing scores (standard IHC practice).
When is a strong PSMD1 DAB signal biologically credible?
Favor staining within intact nuclei or cytoplasm, consistent with the reported tissue pattern, over an isolated membranous rim (HPA tissue IHC; UniProt Q99460 topology). Check cell identity against the section’s morphology: HPA reports high staining in cerebellar Purkinje cells and low staining in lung alveolar cells, so their expected patterns differ (HPA tissue IHC). Discount staining confined to cut edges, folds or necrotic regions, and compare suspicious deposits with a no-primary control (standard IHC practice). Endogenous peroxidase can also produce DAB signal, so confirm that its blocking step worked before interpreting widespread pigment (standard IHC practice). Treat the HPA pattern as supportive evidence rather than definitive validation because its stated reliability remains pending external verification (HPA tissue IHC).
Boster reagents
Best PSMD1 / 26S proteasome non-ATPase regulatory subunit 1 IHC Antibodies
A06293-1 has real IHC data from human paraffin sections (IHC image captions) and IF data from human paraffin sections and A549 cells (IF image captions); catalog reactivity includes human, mouse and rat (catalog).
Real IHC data
Anti-PSMD1 Antibody ®
Cat # A06293-1
A06293-1 has IHC images from human paraffin sections of placenta, bladder cancer, lung cancer and liver cancer (IHC image captions). A06293-1 also has IF images from A549 cells and human paraffin sections of lung, breast and colon cancer (IF image captions).
Which to pick: For tissue IHC, choose A06293-1: its paraffin-section captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (IHC image captions); the fixative is unreported (IHC image captions). For IF/ICC, choose A06293-1 because both applications are listed and its IF images cover A549 cells and human paraffin sections (catalog; IF image captions). For cross-species work, A06293-1 lists human, mouse and rat reactivity (catalog), while the supplied IHC and IF images document human samples only (IHC and IF image captions); clonality is unreported (catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99460 (PSMD1_HUMAN, 26S proteasome non-ATPase regulatory subunit 1).
- Human Protein Atlas. PSMD1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PSMD1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the actin filaments..
- Human Protein Atlas. PSMD1 antibody validation summary (3 antibodies).
- Integrating innovative multiomics and machine learning strategies for prognostic biomarker discovery in hepatocellular carcinoma: guiding personalized treatment strategies with single-cell analysis. Translational cancer research 2026 — PMC13265232.
- Analysis and experimental validation of the innate immune gene PSMD1 in liver hepatocellular carcinoma and pan-cancer. Heliyon 2023 — PMC10623288.
- Exosomal PSMD1 Derived From Dental Pulp Mesenchymal Stem Cells Promotes Microglial M2 Polarization to Alleviate Spinal Cord Injury. CNS neuroscience & therapeutics 2026 — PMC13538453.
- PSMD1 as a prognostic marker and potential target in oropharyngeal cancer. BMC cancer 2023 — PMC10725586.
- PubMed PMID:8816993 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSMD1 (26S proteasome non-ATPase regulatory subunit 1). Expected localisation Cytoplasm and nucleus in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06293-1). Validated IHC antibody: A06293-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Myoepithelial cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A06293-1); 2. Fixation: Image fixative and duration unreported (datasheet A06293-1); 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 A06293-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06293-1); 8. Primary antibody: Rabbit anti-PSMD1, 2-5 μg/ml (datasheet A06293-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06293-1). Watch for: Fixation: Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; no transmembrane segment; epitope coverage unknown (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q99460, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06293-1, PMC13538453, PMC10725586. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMD1/immunohistochemistry-ihc-guide.