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PSMB5 / Proteasome subunit beta type-5 · IHC design guide
Design Immunohistochemistry for PSMB5
Plan chromogenic PSMB5 IHC on paraffin sections using the catalog antibody at 2–5 µg/mL (datasheet A03418-1). Compare nuclear staining across cell types and account for low antibody–RNA concordance when choosing controls and scoring sections (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 tissue staining; cytoplasm is also annotated (HPA tissue IHC; UniProt) | |
| Staining pattern | Variable nuclear staining; high in granular layer and Sertoli cells (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03418-1) | |
| Caveat | Antibody staining has low concordance with tissue RNA (HPA tissue IHC) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; cleavage of residues 1–59 may remove a propeptide epitope (UniProt) |
Section 1
Recommended PSMB5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A03418-1) and one published HCC and adjacent-tissue protocol (PMC9623531) provide starting conditions.
| 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); the published protocol does not specify a retrieval buffer (PMC9623531).
Section 2
What Is the Expected PSMB5 Staining Pattern?
PSMB5 is a proteasome subunit found in the cytoplasm and nucleus, with no transmembrane segment (UniProt P28074). In tissue IHC, expect chiefly nuclear staining in several tissues, including high staining in cerebellar granular-layer cells and testicular Sertoli cells (HPA tissue IHC). HPA rates its tissue pattern Approved but reports low agreement with RNA data and pending external verification (HPA tissue IHC).
| Strong nuclear staining in cerebellar granular-layer cells or testicular Sertoli cells. | This matches HPA's reported high staining in those cell populations and its broader nuclear tissue pattern (HPA tissue IHC). Judge the named cells and their nuclei, rather than treating every cell in the section as an equivalent positive control (HPA tissue IHC). |
| A strong membranous rim or extracellular deposit dominates the slide. | That distribution conflicts with the reported nuclear tissue pattern and PSMB5's cytoplasmic and nuclear locations (HPA tissue IHC; UniProt P28074). Review morphology, control slides and detection conditions before assigning the signal to PSMB5 (general IHC practice). Cytoplasmic staining alone is not a wrong compartment: UniProt also places PSMB5 there (UniProt P28074). |
| Prominent staining appears in adipocytes or appendix glandular cells. | HPA reports PSMB5 as not detected in those specific cell populations (HPA tissue IHC). Consider nonspecific antibody binding or endogenous detection activity, particularly if the signal also appears on a control lacking primary antibody (general IHC practice). HPA's Approved tissue profile still awaits external verification, so a discrepancy needs investigation rather than an automatic biological conclusion (HPA tissue IHC). |
| Haze covers nuclei, stroma and blank spaces with little cell-to-cell distinction. | A diffuse field obscures the cell-specific nuclear pattern reported by HPA (HPA tissue IHC). It can arise from nonspecific binding, residual detection reagent or inadequate washing (general IHC practice). Compare a control lacking primary antibody and inspect whether recognizable granular-layer or Sertoli-cell nuclei remain distinguishable (HPA tissue IHC; general IHC practice). |
| No staining is visible in cerebellar granular-layer cells or testicular Sertoli cells. | These are HPA-reported high-staining cell populations, so an entirely blank result calls for a technical check before scoring the specimen negative (HPA tissue IHC; general IHC practice). Confirm the expected cells are present, then check the catalog antibody's IHC-P instructions, retrieval and detection controls (general IHC practice). |
💡Expected PSMB5 appearanceCall positive a clear, chiefly nuclear chromogenic signal in cerebellar granular-layer cells or testicular Sertoli cells, where HPA reports High staining; a dominant membrane rim or extracellular deposit is discordant (HPA tissue IHC; UniProt P28074).
| Compartment and topology | UniProt places PSMB5 in cytoplasm and nucleus and lists no transmembrane segment; HPA tissue IHC describes nuclear expression in several tissues (UniProt P28074; HPA tissue IHC). Assess nuclear staining first while allowing that cytoplasmic localization is biologically plausible (UniProt P28074). |
| Maturation and epitope interpretation | UniProt annotates a propeptide at residues 1–59 and a mature chain at 60–263 (UniProt P28074). The supplied record does not locate the catalog antibody's epitope, so it cannot establish whether staining recognizes precursor, mature protein or both. |
| Isoforms | UniProt lists 3 isoforms (UniProt P28074). Without an antibody epitope or isoform-specific validation in the supplied evidence, do not assign a nuclear or cytoplasmic signal to a particular isoform. |
| Strength of the tissue reference | HPA labels the tissue IHC pattern Approved, reports low staining-to-RNA consistency and says external verification is pending (HPA tissue IHC). Use its high-staining cells as practical reference points while treating unexpected tissue differences as findings to verify. |
| IF/ICC Q: What location should a separate IF experiment assess? | A: HPA reports supported nucleoplasmic localization, with approved additional centrosome, sperm mid-piece and principal-piece locations (HPA subcellular ICC-IF). Those ICC-IF observations do not establish an IHC-P staining pattern in a new tissue. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected nuclear signal is absent in a high-staining reference tissue. | A missed cell population, unsuitable retrieval or a failed antibody or detection step can produce a blank slide (HPA tissue IHC; general IHC practice). | Locate granular-layer cells or Sertoli cells, verify the run controls, and follow the catalog antibody's IHC-P instructions for retrieval and detection (HPA tissue IHC; general IHC practice). |
| Nuclei stain, but the whole section also has diffuse color. | Nonspecific reagent binding or incomplete washing can reduce contrast (general IHC practice). | Compare the control lacking primary antibody; review blocking, washes and detection conditions while preserving the identifiable nuclear pattern (general IHC practice; HPA tissue IHC). |
| A control lacking primary antibody develops chromogen. | Endogenous detection activity or a detection-reagent artifact may contribute signal independently of primary-antibody binding (general IHC practice). | Check the detection system's relevant endogenous-activity block and reagent controls before interpreting cellular staining (general IHC practice). |
| The strongest signal is on membranes or outside cells. | This is discordant with HPA's nuclear tissue profile and UniProt's cytoplasmic and nuclear annotation (HPA tissue IHC; UniProt P28074). | Inspect section morphology and the control lacking primary antibody, then review staining and detection conditions before scoring PSMB5 (general IHC practice). |
| Adipocytes or appendix glandular cells stain strongly. | Those specific cells are reported as not detected; nonspecific binding or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare staining with a control lacking primary antibody and an HPA-reported high-staining cell population; record unresolved discrepancies (HPA tissue IHC; general IHC practice). |
| A weak slide is being scored as negative throughout. | HPA reports medium staining in several cell populations and low staining in others, so one threshold can obscure differences (HPA tissue IHC). | Score the named cell type and compartment separately, compare with a high-staining reference, and retain HPA's tissue-validation caveat in the interpretation (HPA tissue IHC). |
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 the granular layer). Run adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the cerebellum slide, assess unstained cells or regions as internal background without assuming a particular cell type is PSMB5-negative.
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; an isotype-matched rabbit IgG control matched to the primary antibody’s clonality; and a PSMB5 knockout sample as a biological negative (catalog caption: rabbit primary antibody). For chromogenic detection, quench endogenous peroxidase and check for endogenous biotin background because the reported detection uses a biotinylated secondary, streptavidin–biotin complex, and DAB (catalog caption).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but does not report a fixative; a target-specific fixation window, fixation effect, and dependence on that retrieval condition are unreported (catalog caption: A03418-1). The supplied evidence does not establish whether frozen sections or IF are easier; HPA provides ICC-IF localization images, while the selected-SKU tissue protocol is IHC on paraffin sections (HPA subcellular; catalog caption: A03418-1). In the densely cellular cerebellar granular layer, distinguish cellular signal from diffuse background when scoring (HPA: High in cells in the granular layer; standard IHC practice).
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, cell compartment, detection background and scoring against the supplied IHC evidence.
How should I retrieve PSMB5 when paraffin sections stain weakly?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A03418-1). The reported tissue image used that retrieval, followed by 10% goat serum blocking and 2 µg/ml primary antibody overnight at 4°C; use those conditions as a matched starting point when troubleshooting weak signal (datasheet A03418-1). If staining remains weak, compare a modest increase in heating time on adjacent sections while checking tissue morphology and background, since excessive retrieval can damage sections (standard IHC practice). Include a no-primary control and a consistently processed reference section so stronger DAB deposition is not mistaken for improved PSMB5 detection (standard IHC practice).
Could fixation explain weak or uneven PSMB5 staining?
The selected tissue caption describes a paraffin-embedded section but does not state its fixative, so target-specific PSMB5 fixation sensitivity is unknown (datasheet A03418-1). Record each specimen’s fixative, fixation duration and processing history, then compare sections processed together before changing antibody concentration or retrieval conditions (standard IHC practice). Uneven fixation can alter morphology and antigen accessibility in paraffin sections; inspect the affected area against its hematoxylin counterstain and a consistently processed reference (standard IHC practice). Keep EDTA pH 8.0 retrieval constant during that comparison, because changing fixation history and retrieval together obscures the cause of weak staining (datasheet A03418-1; standard IHC practice).
Should PSMB5 appear in nuclei, cytoplasm, or both?
Evaluate nuclear and cytoplasmic staining separately: PSMB5 is annotated in both compartments, with nuclear translocation following interaction with AKIRIN2 (UniProt P28074 subcellular location). Tissue IHC reports nuclear expression across several tissues, while the subcellular dataset supports nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). A cytoplasmic component is therefore plausible, but score its intensity against the no-primary control and adjacent cells before calling it specific (UniProt P28074 subcellular location; standard IHC practice). Use matched EDTA pH 8.0 retrieval and counterstain across sections when comparing compartments; variable retrieval or dense hematoxylin can distort apparent nuclear-to-cytoplasmic balance (datasheet A03418-1; standard IHC practice).
Can processing or alternative splicing change what this antibody detects?
PSMB5 has 3 annotated isoforms and a precursor propeptide spanning residues 1–59; the mature chain spans residues 60–263 (UniProt P28074 processing and isoforms). The supplied caption does not identify the antibody’s epitope, so staining alone cannot establish whether it recognizes precursor, mature protein, or every isoform (datasheet A03418-1). Check any available epitope documentation before interpreting differences between samples, and compare the same antibody and EDTA pH 8.0 retrieval conditions throughout the series (standard IHC practice; datasheet A03418-1). No transmembrane segment or glycosylation sites are annotated; neither observation resolves antibody isoform specificity or proves that a faint compartment is absent (UniProt P28074 topology and glycosylation).
How can I check a nuclear PSMB5 pattern by multiplex IF?
Treat IF/ICC as a separate assay and validate its antibody conditions independently of the paraffin-section IHC image (datasheet A03418-1; standard IF practice). Pair PSMB5 with a cell-identity marker appropriate to the specimen; a neuronal marker can help identify cerebellar granular-layer cells, which show high tissue-IHC staining (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores, favoring a red or far-red PSMB5 channel when green tissue autofluorescence interferes, and include single-label controls (standard IF practice). PSMB5 has no transmembrane segment and is reported in cytoplasm and nucleus, so use controlled permeabilisation that permits intracellular and nuclear antibody access while preserving morphology (UniProt P28074 topology and subcellular location; standard IF practice).
What should I change when DAB stains the whole section?
First inspect a no-primary control processed through the same biotinylated secondary, streptavidin-biotin complex and DAB steps used in the selected image (datasheet A03418-1; standard IHC practice). If that control stains, check peroxidase blocking, endogenous biotin interference, inadequate washing and nonspecific secondary binding before attributing diffuse brown signal to PSMB5 (standard IHC practice). The image used 10% goat serum blocking and 2 µg/ml primary antibody overnight at 4°C; keep those documented settings visible when adjusting one variable at a time (datasheet A03418-1). Compare background in tissue edges and damaged areas with intact interior cells, because nonspecific deposition can follow section damage (standard IHC practice).
How should I quantify PSMB5 across differently stained sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartment before analysis, then report nuclear and cytoplasmic PSMB5 separately if both are present (UniProt P28074 subcellular location; standard IHC practice). An H-score combines the percentage of cells in each intensity category with its intensity weight; report the percentage positive as well when intensity thresholds could affect comparisons (standard IHC practice). For spatial comparisons, count positive cells per mm² of viable tissue and normalise to the number of eligible cells or measured viable area, as appropriate (standard IHC practice). Hold EDTA pH 8.0 retrieval, DAB development, counterstaining and image-analysis thresholds constant, and exclude folds, edges and necrosis using prespecified rules (datasheet A03418-1; standard IHC practice).
How do I distinguish convincing PSMB5 staining from artefact?
Look for reproducible staining within intact cells and plausible nuclear or cytoplasmic compartments, rather than isolated brown deposits at edges or in necrotic tissue (UniProt P28074 subcellular location; standard IHC practice). High staining in cerebellar granular-layer cells or testicular Sertoli cells can provide context, but a negative field alone does not establish technical failure (HPA tissue IHC). Review the no-primary control for endogenous enzyme or detection-system signal, and compare the DAB pattern with cell boundaries and hematoxylin morphology (standard IHC practice). HPA labels its tissue IHC profile Approved while reporting low consistency with RNA expression and pending external verification, so interpret unexpected cell patterns cautiously (HPA tissue IHC).
Boster reagents
Best PSMB5 / Proteasome subunit beta type-5 IHC Antibodies
A03418-1 has IHC images from human paraffin sections and IF/ICC images from MCF-7 cells; listed reactivity includes human, mouse, and rat (catalog image captions; datasheet: reactivity).
Real IHC data
Anti-PSMB5/MB1 Antibody ®
Cat # A03418-1
A03418-1 shows IHC staining in paraffin sections of human breast cancer, liver cancer, lymphoma, and ovarian serous adenocarcinoma (catalog IHC image captions). A03418-1 also shows IF/ICC staining in MCF-7 cells (catalog IF image caption; datasheet: applications).
Which to pick: Choose A03418-1 for human paraffin-section IHC: its own captions report EDTA retrieval at pH 8.0 and 2 μg/mL primary antibody; the listed IHC range is 2–5 μg/mL (A03418-1 IHC image captions; datasheet: IHC dilution). For IF/ICC, A03418-1 is listed for both applications and shown in MCF-7 cells at 5 μg/mL (datasheet: applications; A03418-1 IF image caption). For cross-species work, A03418-1 lists human, mouse, and rat reactivity, while its IHC images show human tissue only; the fixative is unreported (datasheet: reactivity; A03418-1 IHC 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 tissue staining; cytoplasm is also annotated (HPA tissue IHC; 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: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03418-1); Caveat: Antibody staining has low concordance with tissue RNA (HPA tissue IHC); Isoform / epitope: 3 isoforms; cleavage of residues 1–59 may remove a propeptide epitope (UniProt); Regulation: Low tissue RNA specificity (HPA tissue RNA). Evidence: UniProt P28074, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03418-1, PMC9623531. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMB5/immunohistochemistry-ihc-guide.