This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
PSMC3 / 26S proteasome regulatory subunit 6A · IHC design guide
Design Immunohistochemistry for PSMC3
Plan PSMC3 chromogenic IHC in paraffin sections using the catalog antibody’s documented workflow (datasheet A07208-1). Assess cytoplasmic and nuclear staining (HPA tissue IHC) with cell type, intensity, and control scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSMC3 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 | General cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07208-1) | |
| Positive control | Adipose tissue+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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | No specific expression regulator annotated (UniProt) | |
| Isoform / epitope | No extracellular segment; 0 isoforms, chain 1–439 (UniProt) |
Section 1
Recommended PSMC3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A07208-1). One published PSMC3 protocol for mouse brain paraffin sections follows (PMC12552703: Methods).
| Sample | Paraffin-embedded rat cardiac muscle tissue; fixative not specified (datasheet A07208-1) |
| Fixation | Image fixative and duration unreported (datasheet A07208-1); 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 A07208-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07208-1) |
| Primary antibody | Rabbit anti-PSMC3, 2-5μg/ml (datasheet A07208-1) |
| Primary incubation | Overnight at 4 °C (datasheet A07208-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A07208-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSMC3-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A07208-1), then assess staining and background.
Section 2
What Is the Expected PSMC3 Staining Pattern?
PSMC3 is found in the cytoplasm and nucleus (UniProt P17980), with general cytoplasmic and nuclear tissue staining reported by HPA (HPA tissue IHC). Expect staining in cells such as adipocytes, bronchial respiratory epithelial cells, and bone marrow hematopoietic cells, each reported as High (HPA tissue IHC). HPA rates the tissue pattern Approved, with medium staining–RNA consistency and external verification pending (HPA tissue IHC). PSMC3 has no transmembrane segment (UniProt P17980 topology).
| Cytoplasmic and nuclear chromogenic staining appears in bronchial respiratory epithelial cells. | This fits the reported general tissue pattern and a cell type rated High (HPA tissue IHC). Judge the signal within identifiable cells against the counterstain; staining confined to only one compartment warrants closer review (HPA tissue IHC; general IHC practice). |
| A crisp membrane outline dominates, with little cytoplasmic or nuclear signal. | A dominant membrane pattern conflicts with the reported tissue distribution (HPA tissue IHC) and the absence of a transmembrane segment (UniProt P17980 topology). Check section morphology and controls before interpreting the outline as PSMC3 (general IHC practice). |
| Strong signal appears mainly in a cell population outside the reported High groups. | HPA reports low tissue specificity, so staining elsewhere is not automatically incorrect (HPA tissue IHC). An unexpected cell restricted pattern merits comparison with known positive cells and a negative detection control to assess cross-reactivity or endogenous detection activity (general IHC practice). |
| Color spreads across extracellular spaces or coats many cells without clear boundaries. | That appearance is difficult to assign to PSMC3's reported cellular compartments (HPA tissue IHC; UniProt P17980). Compare a primary antibody omission control and inspect blocking, washes, and chromogen development for nonspecific background (general IHC practice). |
| No signal is visible in bronchial respiratory epithelial cells on a test section. | Those cells are rated High by HPA, making them a useful positive comparison, though HPA's tissue evidence still awaits external verification (HPA tissue IHC). Confirm tissue preservation and detection performance before treating the result as biological absence (general IHC practice). |
💡Expected PSMC3 appearanceA convincing positive shows discernible cytoplasmic and nuclear staining in identifiable cells such as High rated bronchial respiratory epithelium (HPA tissue IHC); an isolated membrane rim or cell free haze calls for control based review (UniProt P17980 topology; general IHC practice).
| Which tissue and cell type is being scored? | Adipocytes, adrenal glandular cells, bone marrow hematopoietic cells, and bronchial respiratory epithelial cells are rated High (HPA tissue IHC). Score the identified cell population rather than assigning one intensity to the whole section (general IHC practice). |
| Does membrane staining fit PSMC3 topology? | PSMC3 has no annotated transmembrane segment and is reported in cytoplasm and nucleus (UniProt P17980). A membrane dominant result needs control review; topology alone cannot identify the source of staining (general IHC practice). |
| How firm is the tissue pattern evidence? | HPA calls tissue IHC Approved but describes medium consistency with RNA data and pending external verification (HPA tissue IHC). HPA006065 is IHC Approved; no Enhanced designation is supplied (HPA antibodies). Use this pattern as a comparison, not proof of antibody specificity. |
| Do processing or modifications predict a separate staining pattern? | UniProt lists one chain spanning residues 1–439, no signal peptide or propeptide, and three modified residues (UniProt P17980). The supplied record gives no antibody epitope, so it cannot establish a modification dependent IHC pattern or retrieval requirement. |
| What should IF/ICC show? | HPA reports mainly nucleoplasm and cytosol, with nuclear bodies as an additional location (HPA subcellular ICC-IF). UniProt also notes colocalization with TRIM5 in cytoplasmic bodies (UniProt P17980). These localization observations guide image interpretation; the IF/ICC workflow belongs in its own guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive comparison tissue has no visible stain. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Check the run's positive control, section morphology, reagent sequence, and chromogen development before interpreting absence in the test tissue (general IHC practice). |
| A membrane rim is stronger than the nuclear and cytoplasmic stain. | Membrane dominant staining does not match HPA's general compartment pattern or UniProt's topology (HPA tissue IHC; UniProt P17980 topology). | Compare the rim with the primary antibody omission control and with nearby clearly identified positive cells; review nonspecific edge staining (general IHC practice). |
| Diffuse color obscures cell boundaries. | Background from detection or incomplete washing can make cellular localization unreadable (general IHC practice). | Inspect a primary antibody omission control, then review blocking, wash steps, and chromogen development time using the same tissue (general IHC practice). |
| Unexpected cells stain while expected cells remain weak. | Low tissue specificity permits broad expression, but a reversed cell pattern also raises a specificity or detection concern (HPA tissue IHC; general IHC practice). | Identify cell types on the counterstained section and compare an HPA High population with a negative detection control before scoring the unexpected signal (HPA tissue IHC; general IHC practice). |
| Nuclear signal is absent although cytoplasmic staining is clear. | The result is narrower than HPA's general cytoplasmic and nuclear tissue profile; staining intensity may vary across cells (HPA tissue IHC). | Recheck nuclear boundaries with the counterstain and compare a positive tissue section from the same run before calling a compartment specific change (general IHC practice). |
| Puncta are scored as background solely because they look unusual. | HPA reports nuclear bodies in ICC-IF, and UniProt reports colocalization with TRIM5 in cytoplasmic bodies (HPA subcellular ICC-IF; UniProt P17980). | First establish whether the surrounding cellular stain and controls support PSMC3; do not infer that every punctum on an IHC section is specific (general IHC practice). |
Experiment evidence
IHC & IF images showing PSMC3 staining
A07208-1 · IHCIHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (A07208-1). TBP-1/PSMC3 was detected in paraffin-embedded section of rat cardiac
A07208-1 · IHCIHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (A07208-1). TBP-1/PSMC3 was detected in paraffin-embedded section of human appen
M07208-2 · IHCIHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (M07208-2). TBP-1/PSMC3 was detected in a paraffin-embedded section of human liv
M07208-2 · IHCIHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (M07208-2). TBP-1/PSMC3 was detected in a paraffin-embedded section of human col
M07208-2 · IFIF analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (M07208-2). TBP-1/PSMC3 was detected in an immunocytochemical section of MCF-7 ce
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000165916-PSMC3/tissue/cerebral+cortex#imid_1957388
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000165916-PSMC3/tissue/colon#imid_1957314
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000165916-PSMC3/tissue/liver#imid_1957307
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000165916-PSMC3/tissue/kidney#imid_1957316
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA006065; green = PSMC3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA006065; green = PSMC3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSMC3 IHC & IF
🧪Start with adipose tissue and confirm staining in adipocytes (HPA: High in adipocytes). HPA detects PSMC3 in all 45 scored tissues, so there is no supported negative tissue or known negative cell population within the positive slide; use no-primary and isotype controls for the negative readout, and investigate any unstained adipocytes as a possible technical failure (HPA: no negative tissue rows; detected in all 45 scored tissues).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: PSMC3 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 PSMC3 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Run secondary-only and matched nonimmune rabbit IgG isotype controls alongside PSMC3-knockout material as a biological specificity control (caption: rabbit primary antibody; standard IHC practice). For the caption’s biotin-based DAB detection, block endogenous peroxidase and check for endogenous biotin signal in adipose tissue (caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07208-1 tissue-IHC caption does not state a fixative (caption: fixative not stated). The demonstrated paraffin-section workflow uses heat retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is required for every specimen (caption: EDTA heat retrieval). HPA ICC-IF images support nucleoplasm and cytosol localization checks in A-431, U-251MG and U2OS, but do not establish that IF or frozen sections are easier; in adipose sections, score the thin cellular rim rather than the empty lipid vacuole (HPA: approved nucleoplasm and cytosol localization; standard histology practice).
HPA tissue IHC evidence for PSMC3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic 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: PSMC3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PSMC3 IHC Tips
Troubleshoot PSMC3 chromogenic IHC in paraffin sections using the catalog antibody’s reported conditions, intracellular localisation, and appropriate staining controls.
What retrieval conditions should I try first for PSMC3 in paraffin sections?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 (datasheet A07208-1). This is the retrieval reported for the catalog antibody’s paraffin section of rat cardiac muscle; the caption does not state a fixative or heating duration (datasheet A07208-1). Keep section thickness, heating, cooling, and antibody incubation consistent while comparing retrieval runs, and include a section processed without primary antibody to assess detection background (standard IHC practice). If staining remains weak, vary heating duration on matched sections before considering another retrieval buffer, and judge improvement by cellular staining and preserved morphology rather than overall DAB darkness (standard IHC practice).
Could fixation explain weak or uneven PSMC3 staining?
Target specific fixation sensitivity is unknown: the catalog antibody’s paraffin tissue caption does not report the fixative (datasheet A07208-1). Record the fixative, fixation duration, tissue processing, and section age for each specimen before attributing a weak result to PSMC3 biology (standard IHC practice). Compare matched sections processed together using the reported EDTA retrieval at pH 8.0 and the same detection reagents, then assess staining alongside tissue morphology (datasheet A07208-1; standard IHC practice). Uneven staining that follows folds, poorly preserved areas, or section edges warrants a processing check; localisation or phosphorylation annotations alone cannot establish a fixation effect (standard IHC practice; UniProt P17980).
Where should convincing PSMC3 staining appear in chromogenic IHC?
Expect intracellular cytoplasmic and nuclear staining, consistent with the reported PSMC3 locations and general tissue IHC profile (UniProt P17980; HPA tissue IHC). The subcellular atlas places PSMC3 mainly in nucleoplasm and cytosol, with additional nuclear body localisation; those IF observations help frame, but do not prove, a paraffin section pattern (HPA subcellular). Score cytoplasmic and nuclear compartments separately when counterstain makes that distinction reliable, and compare the same cell population across sections (standard IHC practice). A membrane only rim is difficult to reconcile with a protein lacking a transmembrane segment and should trigger checks for edge staining, detection background, and antibody specificity (UniProt P17980 topology; standard IHC practice).
How should epitope uncertainty affect interpretation of PSMC3 staining?
The supplied record lists a 439 amino acid PSMC3 chain, no annotated isoforms, and no transmembrane segment (UniProt P17980). It also lists N terminal acetylation and phosphorylation at residues 9 and 376, but the catalog caption does not identify the antibody epitope (UniProt P17980; datasheet A07208-1). Therefore, do not assign a staining difference to an isoform or a particular modification without separate epitope and specificity evidence (standard IHC practice). If retrieval changes the pattern, compare matched sections and cellular compartments under the same detection conditions; a stronger DAB signal alone does not establish which molecular form was detected (standard IHC practice).
How can IF help assess the cellular pattern seen by PSMC3 IHC?
Use IF as a separate localisation check: PSMC3 has approved nucleoplasm and cytosol locations, with additional nuclear body localisation in the subcellular atlas (HPA subcellular). Multiplex PSMC3 with a validated marker for the cell population being assessed, and include single stain controls so overlapping fluorescence can be distinguished from channel bleed through (standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence, favouring a spectral channel with clearer separation from that background (standard IF practice). Because PSMC3 is intracellular and has no transmembrane segment, use a permeabilisation condition that permits access to cytosolic and nuclear epitopes, then assess morphology and background in parallel (UniProt P17980 topology; standard IF practice).
What should I check when DAB staining obscures PSMC3 localisation?
First compare a no primary control with the stained section to identify signal arising from the detection workflow (standard IHC practice). The reported paraffin section used 10% goat serum block, 2 µg/ml primary antibody overnight at 4°C, a biotinylated secondary, and DAB development (datasheet A07208-1). Check endogenous peroxidase blocking, secondary antibody background, and potential endogenous biotin signal when using a biotin based detection system (standard IHC practice). If controls are clean but staining is diffuse, adjust primary incubation or detection development on matched sections and seek a resolved cytoplasmic or nuclear pattern rather than simply reducing all signal (UniProt P17980; standard IHC practice).
How should I quantify PSMC3 chromogenic staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartments before reviewing outcome groups, since PSMC3 can appear in both cytoplasm and nucleus (UniProt P17980; standard IHC practice). For intensity based scoring, record the percentage of cells at each staining intensity and calculate a compartment specific H score; report the percentage positive alongside it (standard IHC practice). If the question concerns abundance of stained cells, count positive cells per mm² of viable tissue and normalise to the corresponding total cells or evaluated area (standard IHC practice). Apply one threshold, counterstain assessment, and exclusion rule for folds or necrosis across slides, with no primary controls informing the background cutoff (standard IHC practice).
When is a PSMC3 positive IHC result biologically credible?
A credible result shows reproducible intracellular staining in identifiable cells, with cytoplasmic or nuclear distribution consistent with PSMC3 localisation (UniProt P17980; HPA tissue IHC). The tissue atlas reports broad expression and high staining in several listed cell populations, but its tissue IHC reliability is Approved with medium consistency and pending external verification (HPA tissue IHC). Treat membrane restricted signal, staining concentrated at section edges or necrotic areas, and signal also present in no primary controls as reasons to investigate artefact (UniProt P17980 topology; standard IHC practice). Review endogenous peroxidase blocking and, for biotin based detection, endogenous biotin background before interpreting DAB intensity as PSMC3 abundance (standard IHC practice).
Boster reagents
Best PSMC3 / 26S proteasome regulatory subunit 6A IHC Antibodies
The catalog shows PSMC3 IHC in human and rat paraffin sections, plus IF/ICC in human MCF-7 cells (catalog IHC and IF captions).
Real IHC data
Anti-TBP-1/PSMC3 Antibody ®
Cat # A07208-1
Real IHC data
Anti-TBP-1/PSMC3 Antibody ® (monoclonal, 4D3)
Cat # M07208-2
A07208-1 has IHC images from rat cardiac muscle and human appendicitis, renal carcinoma, and bladder sections (A07208-1 IHC captions). M07208-2 has IHC images from human liver cancer and colonic adenocarcinoma sections, plus an IF image from MCF-7 cells (M07208-2 IHC and IF captions).
Which to pick: For paraffin-section IHC, choose A07208-1 when a rat tissue example is useful, or M07208-2 for a mouse monoclonal antibody with human tissue examples (A07208-1 IHC caption; M07208-2 catalog: clone 4D3; M07208-2 IHC captions). For IF/ICC, choose M07208-2, which lists both applications and has an MCF-7 IF image (M07208-2 catalog applications and IF caption). Both list human, mouse, and rat reactivity, but the supplied IHC images document human and rat sections only; the captions identify paraffin sections without reporting the fixative (catalog reactivity; A07208-1 and M07208-2 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P17980 (PRS6A_HUMAN, 26S proteasome regulatory subunit 6A).
- Human Protein Atlas. PSMC3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PSMC3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol. In addition localized to the nuclear bodies..
- Human Protein Atlas. PSMC3 antibody validation summary (1 antibodies).
- The impact of PANoptosis-related genes on immune profiles and subtype classification in ischemic stroke. Scientific reports 2025 — PMC12552703.
- Proteasome subunit PSMC3 variants cause neurosensory syndrome combining deafness and cataract due to proteotoxic stress. EMBO molecular medicine 2020 — PMC7338805.
- PubMed PMID:8419915 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSMC3 (26S proteasome regulatory subunit 6A). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07208-1). Validated IHC antibody: A07208-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); 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 rat cardiac muscle tissue; fixative not specified (datasheet A07208-1); 2. Fixation: Image fixative and duration unreported (datasheet A07208-1); 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 A07208-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07208-1); 8. Primary antibody: Rabbit anti-PSMC3, 2-5μg/ml (datasheet A07208-1); 9. Primary incubation: Overnight at 4 °C (datasheet A07208-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: No extracellular segment; 0 isoforms, chain 1–439 (UniProt); Regulation: No specific expression regulator annotated (UniProt). Evidence: UniProt P17980, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol. In addition…, Datasheet A07208-1, PMC12552703. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMC3/immunohistochemistry-ihc-guide.