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PSMA1 / Proteasome subunit alpha type-1 · IHC design guide
Design Immunohistochemistry for PSMA1
Plan chromogenic PSMA1 IHC in paraffin sections using the expected cytoplasmic and nuclear pattern (HPA tissue IHC). This guide covers antibody concentration (datasheet: 2–5 μg/ml), controls and interpretation of tissue staining.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSMA1 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 | Cytoplasm and nucleus (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07591-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07591-1) | |
| Caveat | Smooth muscle cells and chondrocytes show low staining (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms, Short and Long; epitope coverage is unknown (UniProt) |
Section 1
Recommended PSMA1 IHC & IF Protocols
The catalog antibody protocol is accompanied by 2 published PSMA1 IHC protocols for paraffin sections (PMC5790517; PMC9918360).
| Sample | Paraffin-embedded human lung adenocarcinoma tissue; fixative not specified (datasheet A07591-1) |
| Fixation | Image fixative and duration unreported (datasheet A07591-1); 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 A07591-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07591-1) |
| Primary antibody | Rabbit anti-PSMA1, 2-5 μg/ml (datasheet A07591-1) |
| Primary incubation | Overnight at 4 °C (datasheet A07591-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07591-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSMA1-positive staining in glandular cells of adrenal gland (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 retrieval at pH 8.0 (A07591-1 datasheet); the published protocols used Tris/borate/EDTA at pH 8–8.5 (PMC5790517; PMC9918360).
Section 2
What Is the Expected PSMA1 Staining Pattern?
PSMA1 should show cytoplasmic and nuclear staining in tissue sections (HPA: general cytoplasmic and nuclear expression; IHC reliability Supported). High staining is reported in several glandular cell populations, pancreatic exocrine glandular cells, placental cytotrophoblasts, and cerebellar Purkinje cells (HPA: tissue IHC). This intracellular pattern agrees with PSMA1’s cytoplasmic and nuclear localization and lack of a transmembrane segment (UniProt P25786: localization and topology).
| Cytoplasmic and nuclear signal appears in the expected cells, with a readable counterstain. | This matches the tissue pattern (HPA: general cytoplasmic and nuclear expression). High staining in colon glandular cells or pancreatic exocrine glandular cells offers a documented reference for evaluating the run (HPA: High in both populations). Judge distribution within the relevant cells as well as apparent intensity (general IHC practice). |
| Staining is confined to cell membranes, extracellular material, or structures outside the expected cells. | An exclusively surface or extracellular pattern conflicts with the reported intracellular localization (UniProt P25786: cytoplasm, nucleus; no transmembrane segment). Treat it as a suspect result and check controls, morphology, and detection background before assigning PSMA1 positivity (general IHC practice). |
| Strong signal dominates smooth muscle cells or chondrocytes while the expected positive cells are weak. | Those populations have Low staining in the supplied tissue profile (HPA: smooth muscle cells and chondrocytes Low). This mismatch raises concern for cross-reactivity or endogenous detection activity; Low is not a claim of complete absence (HPA: tissue IHC; general IHC practice). |
| Diffuse color obscures cell boundaries or appears similarly across many unrelated structures. | The slide cannot support a compartment-specific interpretation when background overwhelms cellular detail (general IHC practice). Review the no-primary control and detection reagents, then optimize blocking, washes, and antibody concentration according to the assay instructions (general IHC practice). |
| No signal is visible in an expected positive population, such as adrenal glandular cells. | Adrenal glandular cells are reported High, so absence there makes a negative sample result difficult to trust (HPA: adrenal glandular cells High). Check whether morphology is preserved and whether the staining run, retrieval, primary antibody, and detection controls worked (general IHC practice). |
💡Expected PSMA1 appearanceA convincing positive shows cytoplasmic and nuclear staining in the relevant cells, potentially strong in documented High populations (HPA: general pattern and tissue IHC); isolated membrane or extracellular color is suspect (UniProt P25786: intracellular localization and no transmembrane segment).
| Reference tissue and cell population | The tissue profile has low RNA tissue specificity and no listed negative population (HPA: tissue IHC). Use a documented High population as a positive reference; do not turn a Low population into a guaranteed negative control (HPA: tissue IHC). |
| Antibody validation | Four listed antibodies have Supported IHC status, while the supplied list does not mark any as Enhanced (HPA: antibody validation). A Supported designation informs confidence in the reported pattern; it does not establish how a different antibody will perform (HPA: antibody validation; general IHC practice). |
| PSMA1 location and processing | PSMA1 is a 263-residue proteasome alpha subunit reported in cytoplasm and nucleus; it has no annotated signal peptide, propeptide, or transmembrane segment (UniProt P25786). The record alone does not identify an antibody epitope or predict antigen retrieval performance (UniProt P25786: supplied annotations). |
| IF/ICC Q&A: Where should signal appear? | Mainly in the nucleoplasm, with supported nuclear-body localization; centrosome localization is uncertain (HPA: subcellular ICC-IF). This is an IF/ICC localization reference, while the tissue IHC profile reports both cytoplasmic and nuclear staining (HPA: subcellular ICC-IF; tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are unstained. | The run may have inadequate target detection; a blank High reference population conflicts with its reported pattern (HPA: tissue IHC; general IHC practice). | Review the positive control, section quality, retrieval settings, antibody preparation, and detection steps against the assay instructions before interpreting the sample as negative (general IHC practice). |
| Color appears throughout the section, including cell-poor areas. | Widespread color may reflect nonspecific reagent binding or detection background rather than the cellular pattern (general IHC practice; HPA: general cytoplasmic and nuclear expression). | Inspect a no-primary control; review blocking, wash conditions, detection exposure, and antibody concentration without assuming a PSMA1-specific fixation effect (general IHC practice). |
| Signal is strong in smooth muscle cells or chondrocytes. | This differs from their Low levels in the supplied profile, although Low does not mean zero (HPA: tissue IHC). | Compare adjacent expected positive cells and a no-primary control; assess cross-reactivity or endogenous detection activity before calling these cells strongly positive (HPA: tissue IHC; general IHC practice). |
| Only membranes or extracellular spaces stain. | That distribution disagrees with the reported intracellular localization and lack of a transmembrane segment (UniProt P25786). | Check tissue morphology and background controls, then reassess antibody and detection conditions before accepting the localization (general IHC practice). |
| Nuclear signal seems prominent in IF/ICC but less dominant in tissue IHC. | The modalities have different reported summaries: mainly nucleoplasmic in ICC-IF and general cytoplasmic plus nuclear expression in tissue IHC (HPA: subcellular ICC-IF; tissue IHC). | Interpret each image against its own HPA modality reference and confirm that the cell boundaries and nuclear counterstain are readable (HPA: ICC-IF and tissue IHC; general IHC practice). |
| A negative call relies on a smooth muscle or chondrocyte control. | These are listed as Low, and the supplied HPA tissue profile lists no negative population (HPA: tissue IHC). | Use an assay control to evaluate background and a documented High population to confirm detection; describe Low staining as Low rather than absent (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PSMA1 staining
A07591-1 · IHCIHC analysis of PSMA1 using anti-PSMA1 antibody (A07591-1). PSMA1 was detected in a paraffin-embedded section of human lung adenocarcinoma t
A07591-1 · IHCIHC analysis of PSMA1 using anti-PSMA1 antibody (A07591-1). PSMA1 was detected in a paraffin-embedded section of human rectum adenocarcinoma
PB10087 · IHCIHC analysis of PSMA1 using anti-PSMA1 antibody (PB10087). PSMA1 was detected in a paraffin-embedded section of human lung cancer tissue. He
PB10087 · IHCIHC analysis of PSMA1 using anti-PSMA1 antibody (PB10087). PSMA1 was detected in a paraffin-embedded section of human rectum adenocarcinom t
M07617-1 · IHCImmunohistochemical analysis of paraffin-embedded human liver, using KDEL Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000129084-PSMA1/tissue/cerebral+cortex#imid_9452508
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000129084-PSMA1/tissue/colon#imid_9471010
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000129084-PSMA1/tissue/liver#imid_9471011
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000129084-PSMA1/tissue/kidney#imid_9471012
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037646; green = PSMA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037646; green = PSMA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSMA1 IHC & IF
🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells). HPA lists no negative tissue because PSMA1 is detected in all 45 scored tissues, so no-primary and isotype controls carry the negative; cells on the positive slide that show only counterstain may serve as visual comparators, but are not validated PSMA1-negative cells (HPA: no negative rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: PSMA1 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 PSMA1 in A-431, U-251MG, U2OS, PC-3, A-549, HEK293, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and an isotype control matched to the primary antibody’s host species and immunoglobulin class; a PSMA1 knockout specimen or validated peptide block can test biological specificity (standard IHC practice). For chromogenic colon IHC, quench endogenous peroxidase and check background in the control sections (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07591-1 paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). The caption uses heat-mediated EDTA retrieval at pH 8.0, but provides no comparison that establishes retrieval dependency (selected-SKU caption). Frozen-section performance and whether IF is easier are unreported; HPA does show PSMA1 ICC-IF signal mainly in the nucleoplasm, while luminal material in colon should be distinguished from cellular staining during scoring (HPA: subcellular; standard IHC practice).
HPA tissue IHC evidence for PSMA1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PSMA1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PSMA1 IHC Tips
Troubleshoot PSMA1 staining in paraffin sections by checking retrieval, intracellular localisation, controls, and cell-specific scoring.
Which retrieval condition should I try first for weak PSMA1 staining in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A07591-1). The documented tissue-IHC example used that condition before incubating the catalog antibody at 2 μg/ml overnight at 4°C (caption A07591-1). If staining remains weak, compare retrieval duration or a different buffer on adjacent sections while keeping detection and exposure to chromogen constant (standard IHC practice). Assess both nuclear and cytoplasmic staining, since PSMA1 has been reported in both compartments (UniProt P25786; HPA tissue IHC). Include a matched section processed without primary antibody to identify retrieval-associated background (standard IHC practice).
How should I troubleshoot PSMA1 staining when fixation details are missing?
Treat PSMA1-specific sensitivity to fixation as unknown: the selected paraffin-section caption does not report a fixative (caption A07591-1). Record the actual fixative, fixation interval, and tissue processing for each specimen, then compare sections processed alike (standard IHC practice). If signal differs between specimens, first repeat EDTA retrieval at pH 8.0 and the documented 2 μg/ml overnight primary incubation on matched sections (datasheet A07591-1; caption A07591-1). Use an internal positive cell population and a no-primary control to separate lost signal from increased background (standard IHC practice). Neither the tissue staining profile nor PSMA1 topology establishes a target-specific fixation effect (HPA tissue IHC; UniProt P25786).
Should PSMA1 staining be nuclear, cytoplasmic, or both?
Score nuclear and cytoplasmic signal separately because tissue IHC reports general expression in both compartments (HPA tissue IHC). A predominantly nucleoplasmic pattern is also plausible: the supported main location in cellular imaging is nucleoplasm, with nuclear bodies additionally supported (HPA subcellular). UniProt describes cytoplasmic-to-nuclear translocation following interaction with AKIRIN2 and the import receptor IPO9 (UniProt P25786). Compare compartment patterns within the same tissue section, using morphology and a counterstain to define nuclei (standard IHC practice). Treat isolated centrosomal puncta cautiously because that additional location is marked uncertain, and verify any unexpected pattern against controls (HPA subcellular; standard IHC practice).
Can this stain distinguish PSMA1 isoforms or epitope masking?
Do not assign staining to one isoform without antibody epitope information: PSMA1 has Short and Long isoforms, but this payload does not map the catalog antibody's epitope (UniProt P25786; caption A07591-1). The annotated chain spans residues 1–263, and no transmembrane segment is reported (UniProt P25786). Annotated modifications include an N-terminal acetylmethionine and sites at residues 110 and 177; their effect on this antibody's IHC binding is unknown (UniProt P25786). If staining changes with retrieval, compare adjacent sections under identical detection conditions before attributing it to an isoform or modification (standard IHC practice). Record the antibody identity and retrieval condition with each scored specimen (standard IHC practice).
How can IF help investigate an ambiguous PSMA1 IHC pattern?
Use IF as a complementary localisation check, keeping its staining conditions separate from the paraffin-section IHC conditions reported for A07591-1 (caption A07591-1; standard IF practice). In colon, multiplex PSMA1 with an appropriate glandular-cell marker to test the cell assignment, since glandular cells show high tissue-IHC staining there (HPA tissue IHC). Choose a far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and inspect single-label controls for bleed-through (standard IF practice). Permeabilise appropriately to access intracellular PSMA1 epitopes in cytoplasm and nucleus; PSMA1 has no transmembrane segment (UniProt P25786; standard IF practice). Compare nuclear signal with a nuclear counterstain, without treating IF intensity as directly equivalent to chromogenic IHC intensity (standard IF and IHC practice).
What should I check when PSMA1 DAB staining looks diffuse or nonspecific?
First inspect a no-primary section and the tissue edges to distinguish nonspecific detection or edge artefact from cellular staining (standard IHC practice). The documented example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (caption A07591-1). For DAB detection, check the peroxidase block and shorten chromogen development if the control section also darkens (standard IHC practice). Compare nuclear and cytoplasmic staining with preserved cell morphology rather than interpreting diffuse pigment alone as PSMA1 (HPA tissue IHC; standard IHC practice). Keep retrieval at EDTA pH 8.0 while changing one background-control step at a time (datasheet A07591-1; standard IHC practice).
How should I quantify PSMA1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because PSMA1 tissue staining is reported in both cytoplasm and nucleus (HPA tissue IHC). For chromogenic sections, report an H-score from intensity and percentage of positive cells, with nuclear and cytoplasmic scores kept separate (standard IHC practice). Alternatively, report percentage-positive cells or positive-cell density per mm² when the biological question concerns cell prevalence (standard IHC practice). Normalise cell counts to the number of evaluable cells or evaluable tissue area, and exclude folds, edges, and necrotic regions consistently (standard IHC practice). Use identical retrieval, staining, imaging, and threshold rules across compared sections; document the EDTA pH 8.0 retrieval used here (datasheet A07591-1; standard IHC practice).
What makes a PSMA1-positive IHC result convincing rather than artefactual?
A convincing result has cellular nuclear or cytoplasmic staining in well-preserved tissue, consistent with the reported PSMA1 tissue profile (HPA tissue IHC; standard IHC practice). High staining in colon glandular cells or cerebellar Purkinje cells can provide context, while cell identity must still be confirmed morphologically (HPA tissue IHC; standard IHC practice). Treat staining confined to section edges, necrotic areas, or the no-primary control as suspect (standard IHC practice). Diffuse DAB in the control warrants checking endogenous peroxidase blocking and detection reagents before assigning positivity (standard IHC practice). A reproducible pattern in the wrong compartment or cell population needs independent confirmation; nucleoplasmic localisation is supported in cellular imaging (HPA subcellular; standard IHC practice).
Boster reagents
Best PSMA1 / Proteasome subunit alpha type-1 IHC Antibodies
Anti-PSMA1 IHC images cover human lung, rectum, and urothelial cancers and mouse colon (A07591-1/PB10087 image captions); no PSMA1 IF image is supplied (catalog IF images).
Real IHC data
Anti-PSMA1 Antibody ®
Cat # A07591-1
Real IHC data
Anti-Proteasome 20S C2/PSMA1 Antibody ®
Cat # PB10087
Real IHC data
Anti-KDEL Rabbit Monoclonal Antibody
Cat # M07617-1
A07591-1 has paraffin-section IHC images from human lung adenocarcinoma, rectum adenocarcinoma, urothelial carcinoma, and mouse colon; PB10087 has images from human lung cancer and rectum adenocarcinoma (A07591-1/PB10087 image captions). M07617-1 has a human liver paraffin-section IHC image labeled KDEL, not PSMA1 (M07617-1 title/image caption).
Which to pick: For PSMA1 tissue IHC, choose A07591-1 or PB10087: each has its own human paraffin-section image using EDTA pH 8.0 retrieval and 2 μg/ml primary antibody; neither caption reports the fixative (A07591-1/PB10087 image captions). For work across species, A07591-1 has human and mouse IHC images and lists rat reactivity, although rat tissue IHC is unshown (A07591-1 image captions/catalog reactivity). For IF/ICC, M07591 is a rabbit monoclonal listing both applications but has no IF image; exclude M07617-1 from PSMA1 selection because its title and IHC image identify KDEL (M07591 catalog applications/clone/IF images; M07617-1 title/image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P25786 (PSA1_HUMAN, Proteasome subunit alpha type-1).
- Human Protein Atlas. PSMA1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PSMA1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies and centrosome..
- Human Protein Atlas. PSMA1 antibody validation summary (4 antibodies).
- PSMA1, PSMA5, and PSMB2 serve as prognostic biomarkers and are correlated with tumor-infiltrating leukocytes in HCC. Epigenetics 2026 — PMC13577265.
- PSMA1 mediates tumor progression and poor prognosis of gastric carcinoma by deubiquitinating and stabilizing TAZ. Cell death & disease 2022 — PMC9691733.
- Proteomic profiling of antibody-inducing immunogens in tumor tissue identifies PSMA1, LAP3, ANXA3, and maspin as colon cancer markers. Oncotarget 2018 — PMC5790517.
- PSMA1, a Poor Prognostic Factor, Promotes Tumor Growth in Lung Squamous Cell Carcinoma. Disease markers 2023 — PMC9918360.
- PubMed PMID:1888762 — UniProt-cited evidence.
- PubMed PMID:2025653 — UniProt-cited evidence.
- PubMed PMID:1398136 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSMA1 (Proteasome subunit alpha type-1). Expected localisation Cytoplasm and nucleus (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07591-1). Validated IHC antibody: A07591-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung adenocarcinoma tissue; fixative not specified (datasheet A07591-1); 2. Fixation: Image fixative and duration unreported (datasheet A07591-1); 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 A07591-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07591-1); 8. Primary antibody: Rabbit anti-PSMA1, 2-5 μg/ml (datasheet A07591-1); 9. Primary incubation: Overnight at 4 °C (datasheet A07591-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07591-1); Caveat: Smooth muscle cells and chondrocytes show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms, Short and Long; epitope coverage is unknown (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt P25786, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A07591-1, PMC5790517, PMC9918360. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSMA1/immunohistochemistry-ihc-guide.