This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
PSMD3 / 26S proteasome non-ATPase regulatory subunit 3 · IHC design guide
Design Immunohistochemistry for PSMD3
PSMD3 is part of the 19S proteasome regulatory particle (UniProt). This paraffin-section IHC guide covers antibody selection and fixation, with nuclear and cytoplasmic staining expected in most tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSMD3 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 and cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Most tissues: nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| 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 (standard IHC practice; not target-specific); paraformaldehyde-fixed sections shown (datasheet: M08767 IHC-P) (selected-SKU IHC image M08767) | |
| Caveat | Low staining–RNA consistency; verify the pattern (HPA tissue IHC) | |
| Regulation | No specific expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended PSMD3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol appears alongside published PSMD3 IHC methods for breast cancer tissue (PMC6549480) and clinical tissue or xenografts (PMC10278297).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet M08767) |
| Fixation | Image fixative and duration unreported (datasheet M08767); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-PSMD3, 1:250 (datasheet M08767) |
| Primary incubation | 1 hours at room temperature (datasheet M08767) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSMD3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); the breast cancer study used similar retrieval conditions (PMC6549480).
Section 2
What Is the Expected PSMD3 Staining Pattern?
Expect PSMD3 in nuclei and cytoplasm across many tissues, including adrenal glandular cells and bone marrow hematopoietic cells (HPA: tissue IHC). A membrane-only pattern is unexpected because PSMD3 has no transmembrane segment (UniProt O43242: topology). Use these patterns as guides: HPA rates the tissue IHC evidence Approved but reports low agreement between staining and RNA expression, pending external verification (HPA: reliability).
| Nuclear and cytoplasmic staining appears in adrenal glandular cells or bone marrow hematopoietic cells (HPA: High in both). | This matches the reported IHC distribution. Compare signal with the relevant cells in the same section, since HPA reports nuclear and cytoplasmic expression in most tissues rather than identical intensity in every cell (HPA: tissue IHC). |
| Signal outlines cell membranes while nuclei and cytoplasm show little staining. | Treat a membrane-only result as suspect: the reported tissue pattern is nuclear and cytoplasmic (HPA: tissue IHC), and PSMD3 has no transmembrane segment (UniProt O43242: topology). Check morphology and a detection-only control before assigning that signal to PSMD3 (general IHC practice). |
| Skeletal myocytes, adipocytes, or smooth muscle cells stain strongly (HPA: Low in these cells). | This differs from the HPA reference pattern, but Low does not mean absent (HPA: tissue IHC). Compare a known High cell population and a detection-only control; unexpected chromogenic signal can reflect cross-reactivity or endogenous detection activity (general IHC practice). |
| Chromogen covers extracellular spaces or the whole section without clear cellular boundaries. | A noncellular haze cannot establish the reported nuclear and cytoplasmic distribution (HPA: tissue IHC). Review the detection-only control, washing and blocking; widespread background can obscure a real cellular signal (general IHC practice). |
| Adrenal glandular cells remain unstained (HPA: High in adrenal glandular cells). | A blank known-positive region weakens a negative call elsewhere. Verify that the expected cell population is present, then assess the detection control and the catalog antibody's IHC-P conditions (general IHC practice). HPA's Approved rating still carries an external-verification caveat (HPA: reliability). |
💡Expected PSMD3 appearanceCall a result consistent when identifiable cells show nuclear and cytoplasmic staining, especially in HPA High populations; isolated membrane outlines or noncellular haze are suspect (HPA: tissue IHC; UniProt O43242: topology; general IHC practice).
| Reference tissue and cell type (HPA: tissue IHC) | HPA reports High staining in adrenal glandular cells, bone marrow hematopoietic cells and esophageal squamous epithelial cells, but Low staining in skeletal myocytes, adipocytes and smooth muscle cells. Score the relevant cells rather than assigning one intensity to an entire tissue (HPA: tissue IHC; general IHC practice). |
| Subcellular evidence (HPA: tissue IHC and ICC-IF) | Tissue IHC reports nuclear and cytoplasmic staining; ICC-IF places PSMD3 mainly in nucleoplasm, with additional cytosol signal (HPA: tissue IHC; HPA: subcellular). These observations support a compartment check, while UniProt supplies no subcellular annotation (UniProt O43242: record). |
| Antibody validation and evidence limits (HPA: antibodies and reliability) | HPA lists two rabbit antibodies, HPA048972 and CAB017038, as IHC Approved; both are ICC Supported (HPA: antibodies). The tissue profile has low agreement with RNA and awaits external verification, so an Approved label alone does not settle an unexpected pattern (HPA: reliability). |
| Isoforms and unreported epitope (UniProt O43242: isoforms) | UniProt lists two PSMD3 isoforms, but the supplied record gives no antibody epitope or isoform-specific staining evidence (UniProt O43242: isoforms; HPA: antibodies). Do not infer which isoform produces a stained cell or use intensity to separate isoforms. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known High cell population has no chromogenic signal (HPA: tissue IHC). | The target cells may be absent from the section, or an IHC detection step may have failed (general IHC practice). | Confirm cell identity on the counterstain, check a positive section and the detection controls, then compare the run with the catalog antibody's IHC-P instructions (general IHC practice). No PSMD3-specific retrieval condition is supplied here. |
| Cytoplasm stains, but nuclei do not. | The result only partly matches the reported tissue distribution; nuclear and cytoplasmic staining is expected in most tissues (HPA: tissue IHC). | Inspect nuclear detail and compare an HPA High cell population processed in the same run. Interpret a cytoplasm-only result cautiously because HPA's tissue profile awaits external verification (HPA: tissue IHC and reliability; general IHC practice). |
| Membrane outlines dominate the slide. | That pattern conflicts with nuclear and cytoplasmic tissue staining and lacks support from PSMD3 topology (HPA: tissue IHC; UniProt O43242: topology). | Compare cell boundaries with the counterstain and inspect a detection-only control before scoring membrane signal as specific (general IHC practice). |
| A HPA Low population appears as strong as a HPA High population (HPA: tissue IHC). | Cross-reactivity or endogenous chromogenic detection activity is possible; HPA Low is a relative observation, not a negative standard (HPA: tissue IHC; general IHC practice). | Compare both cell populations in matched sections and review detection-only controls. Record the discrepancy rather than forcing a positive or negative designation (general IHC practice; HPA: reliability). |
| Diffuse brown background hides cell boundaries. | Nonspecific reagent binding or endogenous detection activity may contribute to background (general IHC practice). | Examine the detection-only control, blocking and washes, then score PSMD3 only where cellular localisation can be resolved (general IHC practice; HPA: tissue IHC pattern). |
| IF/ICC Q&A: Where should PSMD3 fluorescence appear? | HPA reports mainly nucleoplasmic localisation, with additional cytosol signal in ICC-IF (HPA: subcellular). | Look for nucleoplasmic signal with possible cytosolic signal; use the separate IF/ICC guide for assay design (HPA: subcellular). Do not treat the tissue IHC intensity list as an IF/ICC intensity standard (HPA: tissue IHC and subcellular). |
Experiment evidence
IHC & IF images showing PSMD3 staining
M08767 · IHCM08767 staining PSMD3 in human brain tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Sa
M08767 · IHCM08767 staining PSMD3 in human testis tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). S
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108344-PSMD3/tissue/cerebral+cortex#imid_4303224
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108344-PSMD3/tissue/colon#imid_4303158
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108344-PSMD3/tissue/liver#imid_4303167
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108344-PSMD3/tissue/kidney#imid_4303152
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017038; green = PSMD3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017038; green = PSMD3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSMD3 IHC & IF
🧪Run cerebral cortex first and confirm staining in neuronal cells (High; HPA: cerebral cortex, neuronal cells). HPA detects PSMD3 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat any unstained cells on the positive slide as a local background reference, not a proven PSMD3-negative population (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: PSMD3 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 PSMD3 in A-431, U-251MG, U2OS, HEK293, RPTEC/TERT1, hTERT-RPE1 (serum starved), NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a control immunoglobulin matched to the primary antibody’s host species and isotype, and a PSMD3-knockout biological control where available (standard IHC practice). For chromogenic brain IHC, quench endogenous peroxidase and check for endogenous biotin if using the caption’s biotinylated secondary; assess lipofuscin autofluorescence if examining brain by IF (caption: biotinylated secondary; standard IHC/IF practice).
⚠️Feasibility: A PSMD3-specific fixation window, fixation effect, and antigen-retrieval dependency are unreported in the supplied evidence; optimize retrieval on paraffin sections (supplied target/application evidence). The selected M08767 brain IHC-P caption reports 1/250 primary antibody for 1 hour at room temperature, but its fixative is unreported (M08767 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier; brain lipofuscin can complicate IF interpretation (HPA: nucleoplasm and cytosol in ICC-IF; standard IF practice).
HPA tissue IHC evidence for PSMD3
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PSMD3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PSMD3 IHC Tips
Troubleshoot PSMD3 staining in paraffin sections by checking retrieval, controls, compartment patterns and cell type before comparing chromogenic signal across samples.
How should I adjust retrieval when PSMD3 staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page IHC retrieval setting). Keep section thickness, cooling time and detection conditions constant while comparing retrieval runs, and include a positive control section in each run (standard IHC practice; HPA: high staining in cerebral cortex neuronal cells). If signal remains weak, test a longer citrate incubation on adjacent sections before changing buffer chemistry, because excess heating can damage morphology and increase nonspecific staining (standard IHC practice). Score nuclear and cytoplasmic staining separately when judging improvement (HPA tissue IHC: nuclear and cytoplasmic expression in most tissues).
Could fixation explain weak or uneven PSMD3 staining?
PSMD3 specific sensitivity to fixation is unknown from the supplied evidence, so treat fixation as a variable to document rather than a proven cause (supplied PSMD3 record: no target specific fixation comparison). For paraffin sections, compare blocks with known fixation histories where possible and keep retrieval, antibody incubation and chromogenic development matched (standard IHC practice). Check whether weak staining tracks thick sections, poorly preserved morphology or variable staining across a block, and inspect a control processed alongside the test sections (standard IHC practice). The selected M08767 brain caption reports paraformaldehyde fixed, paraffin embedded sections, but does not establish which fixation conditions best preserve PSMD3 staining (M08767 tissue IHC caption).
Which compartments should show convincing PSMD3 staining?
Assess both nuclei and cytoplasm in tissue sections, since both compartments are reported across most tissues (HPA tissue IHC: nuclear and cytoplasmic expression). A nuclear component is plausible because cell imaging places PSMD3 mainly in the nucleoplasm, with additional cytosolic localisation (HPA subcellular: nucleoplasm and cytosol). Judge the pattern within intact cells against a matched positive control, using the same counterstain and chromogenic development time so faint nuclear signal remains interpretable (standard IHC practice). Predominantly extracellular, vessel lumen or tissue edge colour should prompt a review of section quality and detection background before it is called PSMD3 (standard IHC practice; HPA subcellular: nucleoplasm and cytosol).
How might isoforms or epitope accessibility affect PSMD3 IHC?
PSMD3 has 2 listed isoforms and a PCI domain at residues 286–465, so epitope location matters when interpreting a negative section (UniProt O43242: isoforms and domain). The supplied product caption does not identify the M08767 epitope or establish recognition of both isoforms; check the antibody documentation before making an isoform claim (M08767 tissue IHC caption). Phosphoserines are listed at residues 418 and 430, but their effect on this antibody's staining is unknown (UniProt O43242: modified residues; M08767 tissue IHC caption). If an alternative antibody with a documented, different epitope is available, compare staining on adjacent sections under matched detection conditions (standard IHC practice).
How should I follow up a PSMD3 IHC pattern with multiplex IF?
Treat IF as a separate validation experiment: the selected M08767 evidence describes chromogenic tissue IHC, while cell imaging reports nucleoplasmic and cytosolic PSMD3 (M08767 tissue IHC caption; HPA subcellular). Pair PSMD3 with a validated marker for the cell population under study, and use single stain controls to assess channel bleed through (standard IF practice). Choose fluorophores in cleaner spectral channels after checking the tissue's autofluorescence, especially when signal is faint (standard IF practice). Because PSMD3 has no annotated transmembrane segment and its reported locations are intracellular, assess whether permeabilisation is needed to expose the antibody's epitope in fixed cells (UniProt O43242: topology; HPA subcellular; standard IF practice).
What should I check when brown signal obscures PSMD3 staining?
First inspect a section processed without primary antibody; widespread colour there points to detection chemistry or endogenous activity rather than primary antibody binding (standard chromogenic IHC practice). Include a peroxidase block before HRP based detection, and keep DAB development time consistent across the comparison (standard chromogenic IHC practice). If background appears only with primary antibody, titrate it from the caption's 1:250 incubation condition while holding retrieval and detection constant (M08767 tissue IHC caption; standard IHC practice). Compare the residual pattern with expected nuclear and cytoplasmic staining, and treat edge concentrated colour or deposits outside intact cells cautiously (HPA tissue IHC: nuclear and cytoplasmic expression; standard IHC practice).
How can I compare PSMD3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then record nuclear and cytoplasmic staining separately because both are reported in tissue (HPA tissue IHC: nuclear and cytoplasmic expression; standard IHC practice). Use an H score from staining intensity and the percentage of positive cells, or report positive cell density per mm² when cell abundance is the question (standard IHC practice). Normalise each readout to the number of evaluable cells or the measured viable tissue area, and use identical thresholds, retrieval and DAB development across sections (standard IHC practice). Report compartment specific results and excluded damaged regions so differences in tissue composition do not masquerade as PSMD3 changes (standard IHC practice).
When is an apparent PSMD3 positive signal likely to be artefact?
A credible result shows staining in intact cells with nuclear and/or cytoplasmic distribution, consistent with the reported tissue pattern and cell imaging (HPA tissue IHC: nuclear and cytoplasmic expression; HPA subcellular: nucleoplasm and cytosol). Interpret positivity within the identified cell type: neuronal cells in cerebral cortex and glial cells in caudate have reported high staining, while skeletal muscle myocytes are reported low (HPA tissue IHC: cerebral cortex, caudate and skeletal muscle). Edge concentrated signal, necrotic areas and colour in cell free spaces warrant review before scoring (standard IHC practice). If a no primary control also develops colour, investigate endogenous enzyme activity or detection background (standard chromogenic IHC practice).
Boster reagents
Best PSMD3 / 26S proteasome non-ATPase regulatory subunit 3 IHC Antibodies
M08767 has IHC-P images from human brain and testis paraformaldehyde-fixed paraffin sections (catalog: M08767 image captions). Neither antibody has a listed IF/ICC application or image (catalog: applications and image captions).
Real IHC data
Anti-PSMD3 Antibody (C-Term)
Cat # M08767
M08767 will render with its human brain IHC-P figure (card payload: M08767 image alt). Its catalog also documents human testis IHC-P staining and lists human and mouse reactivity (catalog: M08767 image captions and reactivity).
Which to pick: Choose M08767 for human tissue IHC-P: its images show staining in paraformaldehyde-fixed paraffin brain and testis sections at 1:250 (catalog: M08767 image captions). Neither SKU is listed for IF/ICC, so neither has catalog support as an IF/ICC choice (catalog: applications and image captions). For rat IHC, A08767-1 lists rat reactivity and IHC, but provides no IHC image or reported fixative (catalog: A08767-1 reactivity, applications and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43242 (PSMD3_HUMAN, 26S proteasome non-ATPase regulatory subunit 3).
- Human Protein Atlas. PSMD3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PSMD3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. PSMD3 antibody validation summary (2 antibodies).
- Proteasome 26S Subunit, non-ATPase 3 (PSMD3) Regulates Breast Cancer by Stabilizing HER2 from Degradation. Cancers 2019 — PMC6549480.
- PSMD3-ILF3 signaling cascade drives lung cancer cell proliferation and migration. Biology direct 2023 — PMC10278297.
- Peripheral inflammation preceeding ischemia impairs neuronal survival through mechanisms involving miR-127 in aged animals. Aging cell 2021 — PMC7811844.
- PubMed PMID:9017604 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSMD3 (26S proteasome non-ATPase regulatory subunit 3). Expected localisation Nuclear and cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M08767 at 1:250 (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. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet M08767); 2. Fixation: Image fixative and duration unreported (datasheet M08767); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-PSMD3, 1:250 (datasheet M08767); 9. Primary incubation: 1 hours at room temperature (datasheet M08767). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); paraformaldehyde-fixed sections shown (datasheet: M08767 IHC-P) (selected-SKU IHC image M08767); Caveat: Low staining–RNA consistency; verify the pattern (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: No specific expression regulator annotated (UniProt). Evidence: UniProt O43242, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet M08767, PMC6549480, PMC10278297. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMD3/immunohistochemistry-ihc-guide.