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PSME3 / Proteasome activator complex subunit 3 · IHC design guide
Design Immunohistochemistry for PSME3
Plan paraffin-section PSME3 IHC around ubiquitous nuclear staining (HPA tissue IHC). Start the IHC-validated antibody at 2–5 μg/ml (datasheet A04375-1) and account for possible cytoplasmic localisation in mitotic cells (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSME3 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Nuclear staining in tissue (HPA tissue IHC) | |
| Staining pattern | Ubiquitous nuclear staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04375-1) | |
| Positive control | Adrenal gland+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 fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Mitotic cells may show cytoplasmic localisation (UniProt) | |
| Regulation | Broad tissue expression (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; epitope differences are unreported (UniProt) |
Section 1
Recommended PSME3 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet: A04375-1). Two published PSME3 paraffin-section protocols provide additional starting conditions (PMC11606656; PMC6698586).
| Sample | Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A04375-1) |
| Fixation | Image fixative and duration unreported (datasheet A04375-1); 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 A04375-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04375-1) |
| Primary antibody | Rabbit anti-PSME3, 2-5 μg/ml (datasheet A04375-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04375-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04375-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSME3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A04375-1); consider citrate pH 6.0 as a published alternative (PMC11606656).
Section 2
What Is the Expected PSME3 Staining Pattern?
PSME3 should stain predominantly in nuclei across many cell types in paraffin sections: HPA reports ubiquitous nuclear expression, low tissue specificity, and supported tissue IHC reliability (HPA tissue IHC). High nuclear staining is reported in glandular cells, respiratory epithelial cells, glial cells, and cerebellar granular-layer cells (HPA tissue IHC). A membrane pattern is unexpected because PSME3 has no transmembrane segment (UniProt P61289 topology).
| Clear nuclear chromogen in glandular cells of an HPA-listed tissue, with recognizable tissue structure (HPA tissue IHC). | This fits the reported ubiquitous nuclear pattern; adrenal gland, appendix, breast, and cervix glandular cells are listed as High (HPA tissue IHC). Judge the nuclear pattern alongside a negative detection control before attributing chromogen to PSME3 (standard IHC practice). |
| Predominantly membrane staining, or cytoplasmic staining that overwhelms nuclei in an interphase population. | A dominant membrane pattern conflicts with the lack of a transmembrane segment (UniProt P61289 topology). PSME3 can enter the cytoplasm during mitosis, so occasional cytoplasmic cells need cell-cycle context; broad cytoplasmic staining warrants an artefact or specificity check (UniProt P61289 subcellular location). |
| Strong chromogen in an unexpected cell population while expected nuclei are weak or blank. | HPA reports low tissue specificity, so an unfamiliar cell type alone does not establish cross-reactivity (HPA tissue IHC). A misplaced pattern may reflect off-target binding or endogenous detection activity; compare morphology, the expected nuclear location, and a primary-antibody omission control (standard IHC practice). |
| Diffuse chromogen outside nuclei, including across tissue structures or on areas lacking cells. | Diffuse deposition does not match the reported nuclear IHC profile (HPA tissue IHC). It can arise from nonspecific binding or detection background; assess the primary-antibody omission control and the distribution of chromogen before scoring cells as positive (standard IHC practice). |
| No nuclear signal in an HPA-listed High population, such as bronchial respiratory epithelial cells or caudate glial cells (HPA tissue IHC). | The result conflicts with the reported examples but does not by itself show that PSME3 is absent (HPA tissue IHC). Check section quality, retrieval, antibody dilution, and detection controls as general IHC variables; no PSME3-specific fixation sensitivity is established here (standard IHC practice). |
💡Expected PSME3 appearanceCall positive when distinct nuclear chromogen is present in the relevant cells, with High staining plausible in HPA-listed populations; dominant membrane staining or diffuse cell-free chromogen is suspect (HPA tissue IHC; UniProt P61289 topology; standard IHC practice).
| Cell-cycle location (UniProt P61289 subcellular location). | PSME3 is nuclear and becomes cytoplasmic during mitosis after nuclear-envelope breakdown (UniProt P61289 subcellular location). A few dividing cells with cytoplasmic signal can fit that biology; widespread cytoplasmic IHC signal still needs validation against the predominantly nuclear tissue pattern (HPA tissue IHC). |
| Topology and processing (UniProt P61289 topology and processing). | PSME3 has no transmembrane segment or signal peptide, and its annotated chain spans residues 2–254 (UniProt P61289 topology and processing). This supports an intracellular interpretation; the record supplies no mapped antibody epitope, so it cannot predict retrieval requirements or isoform-specific staining (UniProt P61289 record). |
| Antibody evidence (HPA antibodies; HPA tissue IHC). | The tissue IHC profile has Supported reliability, and both listed HPA antibodies have Supported IHC status (HPA tissue IHC; HPA antibodies). The Enhanced designation shown for HPA012510 applies to ICC, so it should not be transferred to the IHC interpretation (HPA antibodies). |
| IF/ICC Q&A: where should fluorescence appear (HPA subcellular ICC-IF)? | Mainly in the nucleoplasm, a supported location; cytosol, primary cilium, and basal body are additional uncertain locations (HPA subcellular ICC-IF). IF/ICC belongs to its own guide page; these ICC-IF observations do not establish a paraffin IHC protocol or make the uncertain locations required positive criteria (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected High nuclei are blank in a listed tissue (HPA tissue IHC). | Retrieval, staining, or detection may have failed; the supplied record does not identify a PSME3-specific fixation effect (standard IHC practice). | Check tissue integrity and the detection control, then review the antibody's IHC-P instructions and a known-positive section; adjust retrieval or dilution only within a validated IHC workflow (standard IHC practice). |
| Only cytoplasm stains in mostly interphase cells. | That distribution differs from ubiquitous nuclear tissue IHC, although mitotic PSME3 can be cytoplasmic (HPA tissue IHC; UniProt P61289 subcellular location). | Identify mitotic cells morphologically, compare neighboring interphase nuclei, and repeat with appropriate specificity and detection controls before assigning cytoplasmic positivity (standard IHC practice). |
| Membrane outlines dominate the chromogenic image. | A membrane-dominant result conflicts with PSME3's lack of a transmembrane segment (UniProt P61289 topology). | Review section morphology and the primary-antibody omission control; treat persistent membrane staining as unverified until an independently validated IHC antibody supports it (standard IHC practice). |
| Unexpected cell types stain more strongly than expected nuclei. | Low tissue specificity allows broad expression, while cross-reactivity or endogenous detection activity can also produce misleading chromogen (HPA tissue IHC; standard IHC practice). | Compare the cell's compartment with the HPA nuclear pattern and inspect an omission control; confirm a disputed distribution with an independently validated IHC antibody (HPA tissue IHC; standard IHC practice). |
| Chromogen forms a diffuse haze or stains cell-free areas. | Background from nonspecific binding or the detection system can obscure nuclear staining (standard IHC practice). | Inspect the omission control, blocking and wash steps, and chromogen development; rescore only when individual nuclei can be distinguished from background (standard IHC practice). |
| The result looks weaker than an HPA High example. | High is reported for specified tissue and cell populations, while the supplied evidence gives no universal intensity threshold or PSME3-specific fixation rule (HPA tissue IHC). | Compare the same cell type and compartment, review the run's controls, and record the observed intensity without converting an HPA example into a cutoff (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing PSME3 staining
A04375-1 · IHCIHC analysis of PSME3 using anti-PSME3 antibody (A04375-1). PSME3 was detected in a paraffin-embedded section of human cervical cancer tissu
A04375-1 · IHCIHC analysis of PSME3 using anti-PSME3 antibody (A04375-1). PSME3 was detected in a paraffin-embedded section of human colon cancer tissue.
A04375-1 · IFIF analysis of PSME3 using anti-PSME3 antibody (A04375-1) and anti-Tubulin Alpha antibody (M03989-3). PSME3 was detected in immunocytochemic
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000131467-PSME3/tissue/cerebral+cortex#imid_2217172
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000131467-PSME3/tissue/colon#imid_2217082
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000131467-PSME3/tissue/liver#imid_2217099
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000131467-PSME3/tissue/kidney#imid_2217084
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA012510; green = PSME3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA012510; green = PSME3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSME3 IHC & IF
🧪Run appendix first and expect its glandular cells to stain (HPA: High in appendix glandular cells). HPA detects PSME3 in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; any unstained cells within the appendix section should remain at background, but no cell type there is established as PSME3-negative (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: PSME3 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 PSME3 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), KOLF2.1J, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, an isotype control matched to the rabbit primary antibody’s host species and clonality, and a verified PSME3 knockout specimen or validated peptide competition for biological specificity (selected-SKU caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase in appendix inflammatory cells before HRP/DAB detection (selected-SKU caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04375-1 paraffin-section caption does not state a fixative (selected-SKU caption). The caption reports heat retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml overnight at 4°C; it does not establish that retrieval is required (selected-SKU caption). Frozen sections or IF/ICC are not shown to be easier by the supplied evidence; in appendix, peroxidase-positive inflammatory cells can complicate chromogenic interpretation (HPA: High in appendix glandular cells; standard IHC practice).
HPA tissue IHC evidence for PSME3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PSME3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PSME3 IHC Tips
Use nuclear staining as the main readout for PSME3 in chromogenic IHC, while checking retrieval, detection background and cell context (HPA: ubiquitous nuclear expression).
Which retrieval conditions should I start with for PSME3 paraffin sections?
Start with heat-mediated antigen retrieval in EDTA buffer at pH 8.0 (datasheet A04375-1). The selected paraffin-section example used that retrieval before staining human cervical cancer tissue with 2 μg/ml catalog antibody overnight at 4°C (datasheet A04375-1). If nuclear staining is weak, vary heating duration and cooling consistently across a small section series, then compare signal with tissue structure and background (standard IHC practice; HPA: ubiquitous nuclear expression). Record the heating device and duration for each run, because the caption specifies the buffer and pH but gives no retrieval time (datasheet A04375-1).
Could fixation explain weak PSME3 staining in my paraffin sections?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative, and the supplied evidence gives no PSME3 fixation comparison (datasheet A04375-1). Record the fixative, fixation duration and processing history for every specimen before changing the staining protocol (standard IHC practice). Compare sections processed alike, using the documented EDTA pH 8.0 retrieval and 2 μg/ml antibody incubation as a starting point (datasheet A04375-1). If only one processing batch shows weak nuclei, test matched sections with a modest retrieval adjustment and inspect morphology before attributing the difference to fixation (standard IHC practice; HPA: ubiquitous nuclear expression).
How should I assess cytoplasmic PSME3 staining alongside nuclear staining?
Score nuclear staining first, since tissue IHC shows ubiquitous nuclear expression and subcellular data support nucleoplasmic localisation (HPA: ubiquitous nuclear expression; HPA subcellular: nucleoplasm supported). PSME3 can appear in the cytoplasm during mitosis after nuclear envelope breakdown, so note mitotic figures when evaluating cytoplasmic signal (UniProt P61289: cell-cycle localisation). Compare cytoplasmic staining with neighbouring nuclei and with a no-primary control, keeping development conditions identical across sections (standard IHC practice). Diffuse cytoplasmic colour throughout a field without a convincing nuclear pattern warrants a review of background and tissue preservation before biological interpretation (HPA: ubiquitous nuclear expression; standard IHC practice).
Can this antibody distinguish PSME3 isoforms or modification-dependent staining?
PSME3 has 3 annotated isoforms, but the supplied product evidence does not map this antibody’s epitope or establish isoform selectivity (UniProt P61289: isoforms 1, 2 and 3; datasheet A04375-1). The record lists phosphorylation and acetylation sites, including Ser17, Ser24, Lys195 and Ser247, without showing that any changes this antibody’s staining (UniProt P61289: modified residues). Interpret chromogenic signal as PSME3 immunoreactivity rather than a specific isoform or modification state (UniProt P61289: isoforms and modified residues; datasheet A04375-1). For a suspected epitope issue, compare controlled retrieval conditions on adjacent sections and seek direct antibody epitope evidence before assigning a molecular cause (standard IHC practice).
How can I check a PSME3 localisation result by multiplex immunofluorescence?
For a separate IF/ICC experiment, pair PSME3 with a marker identifying the cell population of interest and a nuclear counterstain, then assess signal within individual cells (standard IF practice; HPA subcellular: nucleoplasm supported). Choose a fluorophore channel with low measured tissue autofluorescence and include single-stain controls to check bleed-through (standard IF practice). PSME3 has no annotated transmembrane segment and is mainly nucleoplasmic, so choose permeabilisation that allows antibody access to intracellular epitopes while preserving nuclear structure (UniProt P61289: topology; HPA subcellular: nucleoplasm supported; standard IF practice). Treat additional cytosol, cilium and basal-body signals cautiously because those HPA locations are uncertain (HPA subcellular: additional locations uncertain).
What should I change when brown signal obscures PSME3-positive nuclei?
First compare the section with a no-primary control and inspect whether colour follows tissue edges, damaged areas or the expected nuclei (standard IHC practice; HPA: ubiquitous nuclear expression). For HRP–DAB detection, check the peroxidase block and shorten chromogen development if nonspecific brown colour is widespread (standard IHC practice). The selected example used 10% goat serum blocking, 2 μg/ml antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A04375-1). Change one incubation or detection condition at a time and retain the same counterstain for comparison (standard IHC practice).
How should I quantify PSME3 staining across differently sized tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell populations and analysis regions before scoring, then report the percentage of positive nuclei and an intensity-based H-score for each region (standard IHC practice; HPA: ubiquitous nuclear expression). An H-score combines percentages at intensity levels 0–3 and spans 0–300 (standard IHC practice). For focal populations, report positive nuclei per mm² and normalise counts to the viable analysed area or to the number of eligible cells (standard IHC practice). Keep retrieval, DAB development, imaging and positivity thresholds consistent across sections; report cytoplasmic staining separately because mitotic redistribution can affect its interpretation (standard IHC practice; UniProt P61289: cell-cycle localisation).
What evidence separates true PSME3 staining from a tissue artefact?
A convincing result includes staining in intact nuclei, consistent with ubiquitous nuclear tissue expression and supported nucleoplasmic localisation (HPA: ubiquitous nuclear expression; HPA subcellular: nucleoplasm supported). Check the expected cells within their tissue context, but avoid calling an unstained cell type negative solely because another cell type stains strongly; HPA reports high staining in several different cell populations (HPA: listed tissue IHC positives). Discount edge staining, necrotic regions and signal reproduced in a no-primary control, and investigate endogenous peroxidase when brown colour appears independent of primary antibody (standard IHC practice). Isolated cytoplasmic signal may reflect mitosis, but assess nuclear morphology before making that assignment (UniProt P61289: mitotic cytoplasmic localisation; standard IHC practice).
Boster reagents
Best PSME3 / Proteasome activator complex subunit 3 IHC Antibodies
A04375-1 has IHC images from human cervical and colon cancer paraffin sections and an IF image from U2OS cells (catalog image captions). Catalog reactivity lists human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-PSME3 Antibody ®
Cat # A04375-1
A04375-1 has IHC data from human cervical and colon cancer paraffin sections (A04375-1 IHC image captions). A04375-1 also has IF/ICC data from U2OS cells (A04375-1 IF image caption).
Which to pick: For tissue IHC, choose A04375-1: its own captions document paraffin-section staining after EDTA pH 8.0 retrieval, but do not report the fixative (A04375-1 IHC image captions). For IF/ICC, A04375-1 has a U2OS cell image and an IF/ICC listing (A04375-1 IF image caption; catalog: applications). For cross-species work, A04375-1 is a rabbit polyclonal antibody listed as reactive with human, mouse and rat; the supplied IHC and IF images document human samples (catalog: host, clonality and reactivity; A04375-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P61289 (PSME3_HUMAN, Proteasome activator complex subunit 3).
- Human Protein Atlas. PSME3 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PSME3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol, primary cilium and basal body..
- Human Protein Atlas. PSME3 antibody validation summary (2 antibodies).
- Shikonin Induces Glioma Necroptosis, Stemness Decline, and Impedes (Immuno)Proteasome Activity. Stem cells international 2024 — PMC11606656.
- Downregulated miR-585-3p promotes cell growth and proliferation in colon cancer by upregulating PSME3. OncoTargets and therapy 2019 — PMC6698586.
- PQBP3 prevents senescence by suppressing PSME3-mediated proteasomal Lamin B1 degradation. The EMBO journal 2024 — PMC11405525.
- PSME3 promotes lung adenocarcinoma development by regulating the TGF-β/SMAD signaling pathway. Translational lung cancer research 2024 — PMC11225040.
- PubMed PMID:1968796 — UniProt-cited evidence.
- PubMed PMID:7951316 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSME3 (Proteasome activator complex subunit 3). Expected localisation Nuclear staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04375-1). Validated IHC antibody: A04375-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 cervical cancer tissue; fixative not specified (datasheet A04375-1); 2. Fixation: Image fixative and duration unreported (datasheet A04375-1); 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 A04375-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04375-1); 8. Primary antibody: Rabbit anti-PSME3, 2-5 μg/ml (datasheet A04375-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04375-1). Watch for: Fixation: Keep fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Mitotic cells may show cytoplasmic localisation (UniProt); Isoform / epitope: 3 isoforms; epitope differences are unreported (UniProt); Regulation: Broad tissue expression (HPA tissue IHC). Evidence: UniProt P61289, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A04375-1, PMC11606656, PMC6698586. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSME3/immunohistochemistry-ihc-guide.