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PSMB5 / Proteasome subunit beta type-5 · IHC design guide
Design Immunohistochemistry for PSMB5
Plan PSMB5 paraffin IHC around nuclear staining that varies across tissues (HPA tissue IHC). Use the catalog antibody’s documented IHC conditions (datasheet A03418-1) and include controls because staining has low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSMB5 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 tissues (HPA tissue IHC); cytoplasm also annotated (UniProt) | |
| Staining pattern | Nuclear staining in several tissues at varied intensity (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03418-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 3 isoforms; propeptide 1–59 is removed; check epitope (UniProt) |
Section 1
Recommended PSMB5 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A03418-1) with published PSMB5 staining of piglet brain (PMC10925635) and HCC tissues (PMC9623531).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A03418-1) |
| Fixation | Image fixative and duration unreported (datasheet A03418-1); 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 A03418-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03418-1) |
| Primary antibody | Rabbit anti-PSMB5, 2-5μg/ml (datasheet A03418-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03418-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03418-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSMB5-positive staining in cells in granular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in several tissues with various levels of intensity. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A03418-1); sodium citrate, boiled for 20 min, is a published pig brain alternative (PMC10925635).
Section 2
What Is the Expected PSMB5 Staining Pattern?
In paraffin section IHC, expect PSMB5 mainly in nuclei, with intensity varying by tissue and cell type (HPA: nuclear expression in several tissues). Granular layer cells in cerebellum and Sertoli cells in testis show high staining (HPA: High in both). Cytoplasmic staining is biologically plausible (UniProt P28074: cytoplasm and nucleus; no transmembrane segment). Interpret the pattern cautiously: HPA rates tissue IHC Approved but reports low consistency with RNA expression and pending external verification (HPA: reliability description).
| Strong nuclear staining in cerebellar granular layer cells or testicular Sertoli cells. | This fits the reported high staining in those specific cells (HPA: High in cerebellar granular layer cells; High in Sertoli cells). Compare the expected cell population, compartment and nearby unstained areas before scoring a section positive (general IHC practice). |
| Predominant staining at cell borders, in extracellular material or as an isolated membrane rim. | This conflicts with the reported nuclear tissue pattern and intracellular localization (HPA: nuclear expression; UniProt P28074: cytoplasm and nucleus, no transmembrane segment). Treat a strong border or extracellular pattern as a possible artefact; cytoplasmic signal alone is not automatically wrong (UniProt P28074: cytoplasm). |
| Strong staining in adipocytes or bone marrow hematopoietic cells while expected positive cells stain weakly. | These cell populations are reported as not detected (HPA: adipocytes and bone marrow hematopoietic cells). Consider antibody cross-reactivity or endogenous chromogenic activity, and compare an appropriate negative detection control (general IHC practice). An HPA result for one cell population does not make every cell in that tissue negative. |
| Diffuse color across many cells and tissue spaces, with little nuclear contrast. | A widespread, poorly localized deposit is hard to reconcile with the reported nuclear tissue profile (HPA: nuclear expression in several tissues). Excess detection background, inadequate blocking or insufficient washing can produce this appearance (general IHC practice); evaluate it against a control lacking primary antibody. |
| No signal in cerebellar granular layer cells or testicular Sertoli cells. | Both are reported high staining sites, so a blank result there warrants a technical check (HPA: High in both cell populations). Check section integrity, retrieval, antibody dilution and detection controls as general IHC practice. HPA's Approved tissue rating still carries a low RNA concordance caveat and awaits external verification (HPA: reliability description). |
💡Expected PSMB5 appearanceCall a section positive when the expected cells show discernible, mainly nuclear chromogenic staining—potentially high in cerebellar granular layer cells or Sertoli cells—while a dominant membrane rim or diffuse tissue-wide deposit prompts review (HPA: nuclear expression; High in both cell populations; UniProt P28074: no transmembrane segment).
| Tissue and cell selection | Use the named cell population when judging signal: granular layer cells and Sertoli cells are High, whereas adipocytes are Not detected (HPA: tissue IHC). Low tissue RNA specificity does not predict equal staining in every cell (HPA: low tissue specificity). |
| Location and proteasome biology | UniProt places PSMB5 in cytoplasm and nucleus and describes nuclear import following AKIRIN2 interaction (UniProt P28074: subcellular location). HPA tissue images are summarized as nuclear, so score nuclear and cytoplasmic compartments separately (HPA: tissue IHC profile). |
| Antibody evidence | One listed antibody is IHC Approved; its ICC evidence is Supported (HPA: HPA049518). HPA also reports low consistency between tissue staining and RNA expression, pending external verification (HPA: reliability description). Confirm an unexpected pattern independently before interpreting it as biology (general IHC practice). |
| Processing and isoforms | The precursor has a 1–59 propeptide, a 60–263 mature chain and three annotated isoforms (UniProt P28074: processing; isoforms). Antibody epitope coverage is unspecified here, so these annotations alone cannot explain a weak stain or an isoform-specific pattern. |
| IF/ICC Q&A: should its pattern match IHC exactly? | No exact match is established: ICC-IF is mainly nucleoplasmic, with additional centrosome, sperm mid-piece and principal-piece localization (HPA: subcellular ICC-IF). Tissue IHC is summarized as nuclear (HPA: tissue IHC profile); assess each application against its own evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells are blank. | The reported high-signal cells may have been missed, or a routine IHC step may have failed (HPA: High in cerebellar granular layer cells and Sertoli cells; general IHC practice). | Verify cell identity and section quality, then review retrieval, antibody dilution and detection controls (general IHC practice). No PSMB5-specific retrieval condition or fixation sensitivity is established by the supplied sources. |
| Color is strongest at tissue edges or in empty spaces. | A deposit outside cells does not fit the intracellular locations reported for PSMB5 (UniProt P28074: cytoplasm and nucleus). Uneven reagent coverage or chromogen deposition is a general IHC possibility. | Check reagent coverage, washes and the primary-antibody-omission control; reassess staining within intact cells before assigning a PSMB5 score (general IHC practice). |
| Unexpected cells stain more strongly than expected positive cells. | Cross-reactivity or endogenous detection activity is possible (general IHC practice), especially when the unexpected population is reported Not detected, such as adipocytes (HPA: adipocytes Not detected). | Compare a control lacking primary antibody and, if needed, an independently validated antibody (general IHC practice). Record the exact cell type rather than calling the whole tissue positive or negative. |
| Nuclear and cytoplasmic signals disagree between samples. | Both compartments are annotated, while HPA summarizes tissue IHC as nuclear; compartment balance need not be identical in every observation (UniProt P28074: cytoplasm and nucleus; HPA: nuclear tissue profile). | Score nuclear and cytoplasmic staining separately and compare the same cell type across sections (general IHC practice). Review strong border-only staining as atypical (UniProt P28074: no transmembrane segment). |
| Most of the section has diffuse chromogenic background. | Blocking, washing or endogenous detection activity can obscure localized staining (general IHC practice). HPA's nuclear profile cannot by itself identify which workflow step caused background (HPA: tissue IHC profile). | Inspect the primary-antibody-omission control, then review blocking, washes, detection reagents and chromogen development (general IHC practice). Judge positivity only where cellular boundaries and nuclear contrast remain interpretable. |
| IHC and IF/ICC show different additional structures. | ICC-IF lists centrosome and sperm-piece locations in addition to nucleoplasm, whereas the tissue IHC summary emphasizes nuclei (HPA: subcellular ICC-IF; tissue IHC profile). | Compare each result with evidence for its own application and cell type; do not use an ICC-IF feature alone to score a paraffin section positive (HPA: subcellular ICC-IF; tissue IHC profile). |
Experiment evidence
IHC & IF images showing PSMB5 staining
A03418-1 · IHCIHC analysis of PSMB5/MB1 using anti-PSMB5/MB1 antibody (A03418-1). PSMB5/MB1 was detected in paraffin-embedded section of human breast canc
A03418-1 · IHCIHC analysis of PSMB5/MB1 using anti-PSMB5/MB1 antibody (A03418-1). PSMB5/MB1 was detected in paraffin-embedded section of human liver cance
A03418-1 · IFIF analysis of PSMB5/MB1 using anti-PSMB5/MB1 antibody (A03418-1). PSMB5/MB1 was detected in immunocytochemical section of MCF-7 cells. Enzy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100804-PSMB5/tissue/cerebral+cortex#imid_13709969
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000100804-PSMB5/tissue/cerebellum#imid_13709972
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100804-PSMB5/tissue/colon#imid_13709031
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100804-PSMB5/tissue/liver#imid_13709024
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA049518; green = PSMB5, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA049518; green = PSMB5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSMB5 IHC & IF
🧪Run cerebellum first and require staining in cells of the granular layer (HPA: High in cells in granular layer); use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes). On the cerebellum slide, genuinely negative cells should show counterstain without DAB signal, but the supplied HPA rows do not identify a validated negative cell type within that tissue.
Positive control tissue: Cerebellum (Cells in granular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PSMB5 in Hep-G2, PC-3, U2OS, Sperm, NIH 3T3, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (hero caption: rabbit primary antibody). Use PSMB5-knockout material, if available, as a biological specificity control; check for endogenous peroxidase and biotin background because the caption uses SABC with DAB (hero caption: SABC/DAB detection).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A03418-1 caption does not state a fixative (selected-SKU caption: fixative not stated). The demonstrated paraffin-section procedure used heat-mediated EDTA retrieval at pH 8.0; whether retrieval is essential, or frozen sections or IF are easier, is unreported (selected-SKU caption: EDTA retrieval; supplied evidence: no matched comparison). In cerebellum, assess DAB background in the densely packed granular layer alongside the controls (HPA: High in cells in granular layer).
HPA tissue IHC evidence for PSMB5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Testis | Sertoli cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PSMB5 IHC Tips
Troubleshoot PSMB5 staining in paraffin sections by checking retrieval, controls, compartment-specific signal, and reproducible scoring (datasheet A03418-1; UniProt P28074 localisation).
How should I adjust retrieval when PSMB5 staining is weak or tissue morphology deteriorates?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03418-1). If staining is weak, compare a longer heating interval or citrate at pH 6.0 on adjacent sections while holding detection conditions constant (standard IHC practice). Use 2 µg/mL primary antibody overnight at 4°C as the documented starting condition, so an incubation change does not confound the retrieval comparison (datasheet A03418-1). If sections detach or morphology deteriorates, shorten heating and compare staining with a no-primary slide to assess retrieval-related background (standard IHC practice).
Could fixation explain inconsistent PSMB5 staining between paraffin specimens?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity for PSMB5 is unknown (datasheet A03418-1). Record each specimen’s fixative and fixation duration, then compare matched sections using EDTA pH 8.0 retrieval and the documented 2 µg/mL primary concentration (datasheet A03418-1; standard IHC practice). If a specimen stains weakly, check a concurrently processed positive section before attributing the result to fixation (standard IHC practice). Assess nuclear and cytoplasmic staining separately, and use preserved morphology plus a no-primary control to help identify nonspecific deposits (UniProt P28074 localisation; standard IHC practice).
Should PSMB5 appear in nuclei, cytoplasm, or both?
Assess both nuclear and cytoplasmic compartments: PSMB5 is annotated in each, with nuclear translocation linked to AKIRIN2 and IPO9 (UniProt P28074 localisation). Tissue IHC reports nuclear expression in several tissues, while subcellular imaging supports a mainly nucleoplasmic location (HPA tissue IHC; HPA subcellular). In the documented paraffin-section workflow, score convincing intracellular staining against nearby background and a no-primary control rather than requiring every cell to show identical compartment intensity (datasheet A03418-1; standard IHC practice). If staining outlines cell membranes exclusively, inspect counterstained morphology and detection controls before calling it PSMB5, which has no transmembrane segment (UniProt P28074 topology; standard IHC practice).
Could propeptide processing or isoforms change what the antibody detects in sections?
PSMB5 has 3 annotated isoforms, and its 1–59 propeptide is removed to yield the 60–263 mature chain (UniProt P28074 processing and isoforms). The supplied antibody caption gives no epitope sequence, so it cannot establish which isoforms or processing states the catalog antibody detects (datasheet A03418-1). Check the antibody’s documented immunogen or epitope against those sequences before interpreting a staining difference as isoform-specific expression (UniProt P28074 processing and isoforms; standard IHC practice). Compare sections under identical EDTA pH 8.0 retrieval and detection conditions; a staining pattern alone cannot resolve precursor from mature PSMB5 (datasheet A03418-1; standard IHC practice).
How can IF help investigate an ambiguous PSMB5 IHC pattern?
Use IF as a follow-up to the paraffin-section IHC pattern, and multiplex PSMB5 with a separately validated cell-type marker where identity is uncertain (standard IF practice). For example, HPA reports high tissue-IHC staining in cerebellar granular-layer cells and testis Sertoli cells, which can guide marker selection (HPA tissue IHC). Choose spectrally separated fluorophores after checking tissue autofluorescence, especially if a weak channel overlaps the tissue’s intrinsic emission (standard IF practice). PSMB5 is intracellular with no transmembrane segment, so permeabilisation must permit access to its nuclear or cytoplasmic epitope; the antibody’s epitope and IF fixation requirements are unspecified here (UniProt P28074 topology and localisation; datasheet A03418-1).
How do I distinguish PSMB5 staining from chromogenic detection background?
The documented workflow uses 10% goat serum blocking, a biotinylated secondary antibody, a streptavidin–biotin complex, and DAB (datasheet A03418-1). Run a no-primary section to reveal secondary or detection-system deposits, and include a peroxidase block before DAB development (standard IHC practice). If diffuse signal persists, check endogenous biotin and compare an avidin–biotin blocking step with the original conditions before changing the PSMB5 antibody concentration (standard IHC practice). Inspect tissue edges and damaged areas separately from intact cells; pigment or precipitate without plausible intracellular localisation should not be scored as PSMB5 (UniProt P28074 localisation; standard IHC practice).
What is a defensible way to quantify PSMB5 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartments before scoring, because PSMB5 can appear in nuclei and cytoplasm (UniProt P28074 localisation). Report the percentage of positive cells and an intensity-based H-score for each compartment, using the same thresholds and exposure to DAB development across sections (standard IHC practice). Normalise positive counts to the number of evaluable cells, or positive-cell density to the measured tissue area in mm², while excluding necrotic and detached regions (standard IHC practice). Include a concurrently stained reference section and record retrieval and antibody concentration, since the documented starting conditions are EDTA pH 8.0 and 2 µg/mL (datasheet A03418-1; standard IHC practice).
When is a PSMB5-positive IHC pattern credible rather than artefactual?
A credible result shows reproducible intracellular staining in intact cells, with nuclear or cytoplasmic signal consistent with PSMB5 localisation (UniProt P28074 localisation; standard IHC practice). Compare cell identity and compartment with HPA’s nuclear tissue profile, but treat that reference cautiously because its antibody staining has low consistency with RNA expression (HPA tissue IHC). Reject isolated staining at cut edges, within necrosis, or in a compartment that cannot be reconciled with cellular morphology until controls resolve it (standard IHC practice). A no-primary section and peroxidase-blocked DAB workflow help identify detection deposits or endogenous enzyme activity before positive cells are scored (standard IHC practice).
Boster reagents
Best PSMB5 / Proteasome subunit beta type-5 IHC Antibodies
A03418-1 has real IHC data from human paraffin sections and IF data from MCF-7 cells (catalog image captions). Its listed reactivity is human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-PSMB5/MB1 Antibody ®
Cat # A03418-1
A03418-1 is listed for IHC and IF/ICC (catalog: applications). Its images show IHC in human breast cancer, liver cancer, lymphoma and ovarian serous adenocarcinoma paraffin sections, plus IF in MCF-7 cells (catalog image captions).
Which to pick: For tissue IHC, choose A03418-1: its own IHC captions show staining in human paraffin sections; the fixative is unreported (catalog IHC image captions). For IF/ICC, the same SKU has an MCF-7 IF image and lists both applications (catalog IF image caption; catalog: applications). It lists human, mouse and rat reactivity, but the supplied IHC and IF images demonstrate human samples only (catalog: reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P28074 (PSB5_HUMAN, Proteasome subunit beta type-5).
- Human Protein Atlas. PSMB5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PSMB5 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the centrosome, mid piece and principal piece..
- Human Protein Atlas. PSMB5 antibody validation summary (2 antibodies).
- Ubiquitination and degradation of CD47 enhances macrophage phagocytosis of hemolytic erythrocytes. iScience 2026 — PMC12818076.
- Proteasome localization and activity in pig brain and in vivo small molecule screening for activators. Frontiers in cellular neuroscience 2024 — PMC10925635.
- A Four-Cell-Senescence-Regulator-Gene Prognostic Index Verified by Genome-Wide CRISPR Can Depict the Tumor Microenvironment and Guide Clinical Treatment of Bladder Cancer. Frontiers in immunology 2022 — PMC9312376.
- PSMB5 overexpression is correlated with tumor proliferation and poor prognosis in hepatocellular carcinoma. FEBS open bio 2022 — PMC9623531.
- PubMed PMID:8753855 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSMB5 (Proteasome subunit beta type-5). Expected localisation Nuclear staining in tissues (HPA tissue IHC); cytoplasm also annotated (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03418-1). Validated IHC antibody: A03418-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in granular layer); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A03418-1); 2. Fixation: Image fixative and duration unreported (datasheet A03418-1); 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 A03418-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03418-1); 8. Primary antibody: Rabbit anti-PSMB5, 2-5μg/ml (datasheet A03418-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03418-1). Watch for: Fixation: Keep fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; propeptide 1–59 is removed; check epitope (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt P28074, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03418-1, PMC10925635, PMC9623531. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMB5/immunohistochemistry-ihc-guide.