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PSPC1 / Paraspeckle component 1 · IHC design guide
Design Immunohistochemistry for PSPC1
Plan chromogenic IHC for PSPC1 in paraffin sections using its predominantly nuclear tissue staining as the expected pattern (HPA tissue IHC). The guide covers fixation consistency, interpretation across cell types, and the limits of the available isoform evidence (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PSPC1 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 | Predominantly nuclear in tissue sections (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across a majority of tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06348-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 44 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Ovarian stromal cells show low staining (HPA tissue IHC) | |
| Regulation | No intensity regulator specified (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended PSPC1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A06348-1) is accompanied by four published PSPC1 IHC methods (PMC4395133; PMC8053714; PMC9184599; PMC6914800).
| Sample | Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A06348-1) |
| Fixation | Image fixative and duration unreported (datasheet A06348-1); 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 A06348-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06348-1) |
| Primary antibody | Rabbit anti-PSPC1, 2-5 μg/ml (datasheet A06348-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06348-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06348-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PSPC1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in a majority of tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A06348-1). The published methods provide glycine, EDTA pH 9.0, and citrate alternatives (cited PMC methods).
Section 2
What Is the Expected PSPC1 Staining Pattern?
PSPC1 is primarily nuclear, including paraspeckles and the nucleolus, with cytoplasmic localization also reported (UniProt Q8WXF1). In paraffin-section IHC, expect nuclear staining across many tissues; HPA reports high staining in selected glandular, endothelial, glial and stromal cells (HPA tissue IHC: Supported; nuclear expression in a majority of tissues). PSPC1 has no transmembrane segment (UniProt Q8WXF1 topology).
| Distinct nuclear chromogen in appendix or duodenum glandular cells. | This fits reported high staining in those cells (HPA tissue IHC: High in appendix and duodenum glandular cells). Judge signal against nearby nuclei and the negative control; a uniformly colored section is less convincing than cell-resolved staining (general IHC practice). |
| Nuclear staining appears punctate, with some nucleolar association. | This is compatible with PSPC1 in paraspeckles and the nucleolus (UniProt Q8WXF1 localization). Routine chromogenic IHC may not resolve individual paraspeckles, so puncta are supportive rather than required (general IHC interpretation). |
| The dominant stain outlines cell membranes or fills extracellular space instead of nuclei. | That distribution conflicts with the mainly nuclear tissue pattern (HPA tissue IHC) and the absence of a transmembrane segment (UniProt Q8WXF1 topology). Consider nonspecific binding, precipitate or detection artefact; compare morphology and control sections (general IHC practice). |
| Strong staining appears in unexpected cells while expected nuclei remain weak. | Cell-specific intensity can vary, but an inverted pattern relative to the selected reference tissue warrants scrutiny (HPA tissue IHC: listed high and low cell groups). Cross-reactivity or endogenous detection activity are possibilities, not diagnoses; assess controls before assigning PSPC1 positivity (general IHC practice). |
| A reference section shows no convincing nuclear signal. | A negative result in appendix or duodenum glandular cells disagrees with their reported high staining (HPA tissue IHC). First check whether the cells are present and the detection run worked; this single section cannot establish target absence (general IHC practice). |
💡Expected PSPC1 appearanceCall the stain positive when cell-resolved nuclear signal is clear in an HPA high cell group, such as duodenum glandular cells (HPA tissue IHC: High); treat dominant membranous or extracellular color as suspect against the nuclear reference pattern (HPA tissue IHC; UniProt Q8WXF1 topology).
| Reference-cell choice | Appendix and duodenum glandular cells, caudate glia, and cerebral-cortex or colon endothelium are reported high (HPA tissue IHC). Cardiomyocytes and ovarian stromal cells are reported low (HPA tissue IHC); a weak result there is less informative as a run check. |
| Subnuclear resolution | PSPC1 occupies paraspeckles and can associate with the nucleolus (UniProt Q8WXF1 localization). HPA tissue IHC reports nuclear expression broadly, without requiring visible puncta in each chromogenic section; score the nuclear compartment before interpreting fine texture. |
| Antibody evidence | The tissue pattern has Supported reliability with high agreement between staining and RNA data (HPA tissue IHC). HPA038904 is listed as Supported for IHC (HPA antibodies); that status supports use as a reference, while a control section still checks each staining run (general IHC practice). |
| Protein forms and epitope limits | UniProt lists two isoforms and a full-length chain spanning residues 1–523, with no signal peptide or propeptide (UniProt Q8WXF1). The supplied record gives no antibody epitope, so isoform-specific detection or epitope-dependent retrieval cannot be predicted from it. |
| IF/ICC Q&A: what pattern should IF show? | HPA reports mainly nucleoplasmic signal with additional nucleolar fibrillar-center localization, and lists A-431, U-251MG, U2OS and HEK293 images (HPA subcellular ICC-IF). Use the separate IF/ICC guide for that application; these cell images do not set an IHC scoring threshold. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in a section selected as a positive reference. | The sampled field may lack the expected cells, or the IHC run may have failed (general IHC practice). Appendix glandular cells are reported high (HPA tissue IHC). | Locate intact glandular cells, inspect counterstain and tissue preservation, then review the run control and detection steps (general IHC practice). |
| Nuclei are weak, but broad brown haze covers the section. | Background can obscure a nuclear pattern through nonspecific binding or incomplete washing (general IHC practice); HPA describes predominantly nuclear tissue expression (HPA tissue IHC). | Compare the no-primary control, wash and blocking steps, and antibody dilution; retain a setting that reveals cell-resolved nuclei (general IHC practice). |
| Cytoplasm is darker than nuclei throughout the reference tissue. | UniProt also reports cytoplasmic localization, but widespread cytoplasm-dominant IHC conflicts with HPA's nuclear tissue profile (UniProt Q8WXF1; HPA tissue IHC). | Review cell boundaries and the no-primary control; repeat with the IHC-validated antibody conditions before interpreting cytoplasmic signal (general IHC practice). |
| Brown signal persists without primary antibody. | Endogenous detection activity or nonspecific detection reagents can produce color in chromogenic IHC (general IHC practice). Such signal cannot establish PSPC1 localization. | Check the detection chemistry and its blocking controls; compare the affected cells with the primary-antibody section (general IHC practice). |
| A low-reference cell group stains strongly. | Cardiomyocytes and ovarian stromal cells are listed as low, so strong staining merits review, although HPA levels do not define an absolute negative control (HPA tissue IHC). | Compare nuclear location and intensity with an HPA high cell group on the same run; inspect no-primary staining before considering cross-reactivity (general IHC practice). |
| IF images show fine puncta, but the IHC section shows smooth nuclear color. | Chromogenic tissue sections may not resolve the structures visible in ICC-IF (general imaging practice). HPA reports nucleoplasmic and nucleolar fibrillar-center IF signal (HPA subcellular ICC-IF). | Score IHC by convincing nuclear, cell-resolved staining and the chosen tissue reference; do not require individually resolved paraspeckles (HPA tissue IHC; UniProt Q8WXF1 localization). |
Experiment evidence
IHC & IF images showing PSPC1 staining
A06348-1 · IHCIHC analysis of PSPC1 using anti-PSPC1 antibody (A16132-1). PSPC1 was detected in a paraffin-embedded section of human glioblastoma tissue.
A06348-1 · IHCIHC analysis of PSPC1 using anti-PSPC1 antibody (A16132-1). PSPC1 was detected in a paraffin-embedded section of human ovarian serous adenoc
A06348-1 · IFIF analysis of PSPC1 using anti-PSPC1 antibody (A06348-1) and anti-Beta Tubulin antibody (M01857-3). PSPC1 was detected in immunocytochemica
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000121390-PSPC1/tissue/cerebral+cortex#imid_9900172
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000121390-PSPC1/tissue/colon#imid_9908050
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000121390-PSPC1/tissue/liver#imid_9908051
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000121390-PSPC1/tissue/pancreas#imid_9908002
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038904; green = PSPC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038904; green = PSPC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PSPC1 IHC & IF
🧪Run appendix first; glandular cells should stain (HPA: High in appendix glandular cells). HPA detects PSPC1 in all 44 scored tissues, so use no-primary and isotype controls for the negative; assess unstained areas or cells on the appendix slide as background only, not as confirmed PSPC1-negative cells (HPA: no negative tissue rows).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: PSPC1 is detected in all 44 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 PSPC1 in A-431, U-251MG, U2OS, HEK293, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Run a secondary-only slide and a concentration-matched rabbit IgG isotype control; use matched PSPC1-knockout material as a biological specificity control (caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection and check for residual tissue background (caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected tissue-IHC caption does not state a fixative (selected tissue-IHC caption). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected tissue-IHC caption). HPA shows ICC-IF images in A-431, U-251MG, U2OS and HEK293 cells; the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC, while HRP/DAB staining warrants a check for endogenous peroxidase background (HPA: subcellular ICC-IF images; caption: HRP/DAB detection; standard IHC practice).
HPA tissue IHC evidence for PSPC1
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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PSPC1 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PSPC1 IHC Tips
Use nuclear staining and cell type controls to assess PSPC1 in chromogenic IHC of paraffin sections (HPA tissue IHC; UniProt Q8WXF1).
Which retrieval condition should I start with for PSPC1 staining in paraffin sections?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 for this page’s IHC assay (datasheet A06348-1). Keep heating, cooling and wash conditions consistent across sections so differences in nuclear staining are interpretable (standard IHC practice). If staining remains weak, adjust heating time on matched sections before treating a different buffer as an experimental fallback (standard IHC practice). Assess signal in viable nuclei and compare it with a no primary control, since PSPC1 is mainly nuclear in tissue IHC (HPA tissue IHC; standard IHC practice). Record any retrieval change alongside antibody identity, because the supplied tissue caption names A16132-1 rather than A06348-1 (selected tissue IHC caption).
How should I troubleshoot weak PSPC1 staining when section fixation history varies?
Target specific sensitivity to fixation is unknown from the supplied evidence, so record the fixative and processing history for each paraffin section (supplied evidence; standard IHC practice). Compare sections with similar handling before attributing weak nuclear signal to fixation, since delayed or prolonged fixation can change antigen accessibility in IHC generally (standard IHC practice). Keep the EDTA pH 8.0 retrieval condition fixed while assessing that possibility (datasheet A06348-1; standard IHC practice). Examine morphology and the no primary control alongside staining; damaged tissue or nonspecific DAB can obscure nuclear scoring (standard IHC practice). The tissue IHC caption reports paraffin embedding but does not state a fixative, so it cannot establish a PSPC1 fixation preference (selected tissue IHC caption).
What staining pattern should I expect, and when is cytoplasmic signal concerning?
Expect predominantly nuclear staining across many tissues, consistent with the supported tissue IHC profile (HPA tissue IHC). PSPC1 is mainly nucleoplasmic in cell imaging, with additional nucleolar fibrillar center localisation; UniProt also describes paraspeckles, nucleolus, nuclear matrix and cytoplasm (HPA subcellular; UniProt Q8WXF1). Routine chromogenic sections may not resolve individual paraspeckles, so score the nuclear compartment before assigning a punctate subnuclear pattern (standard IHC practice). Cytoplasmic staining alone warrants checking the no primary control and tissue morphology, although cytoplasmic localisation is documented and cannot be ruled out categorically (UniProt Q8WXF1; standard IHC practice). Compare cell types: glandular cells in appendix stain high, while cardiomyocytes in heart muscle are reported low (HPA tissue IHC).
How can isoforms or epitope accessibility affect interpretation of the nuclear signal?
PSPC1 has 2 listed isoforms, but the supplied antibody evidence does not identify which isoform its epitope detects (UniProt Q8WXF1; supplied evidence). Its 2 RNA recognition motifs span residues 82–154 and 156–237, making antibody epitope mapping relevant when comparing reagents (UniProt Q8WXF1). Reported modified residues include phosphoserines at 409, 473 and 477, without evidence here that they change this antibody’s staining (UniProt Q8WXF1; supplied evidence). If results differ between antibodies, compare documented immunogen regions and test each under the same retrieval and detection conditions (standard IHC practice). Do not assign a particular isoform or modification from DAB intensity alone; this assay does not provide that molecular resolution (standard IHC practice).
How should I plan a confirmatory PSPC1 immunofluorescence experiment?
For IF/ICC confirmation, multiplex PSPC1 with an independently validated marker of the expected cell type; glial cells in caudate are one high staining reference (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and assess unstained material for tissue autofluorescence before assigning faint puncta to PSPC1 (standard IF practice). Because PSPC1 is mainly nucleoplasmic and has no transmembrane segment, use permeabilisation appropriate for access to a nuclear epitope (HPA subcellular; UniProt Q8WXF1; standard IF practice). Include single stain and no primary controls when judging colocalisation and bleed through (standard IF practice). The supplied IHC caption does not establish IF fixation, permeabilisation or antibody dilution conditions (selected tissue IHC caption).
What controls help distinguish PSPC1 staining from chromogenic background?
Run a no primary control through the same secondary antibody and DAB steps to reveal detection related background (standard IHC practice). Apply a peroxidase block and inspect blood rich or damaged areas for residual enzyme driven colour before calling cells positive (standard IHC practice). Check whether apparent nuclear staining follows intact nuclei rather than folds, section edges or necrotic debris (standard IHC practice). Compare high staining appendix glandular cells with low staining heart muscle cardiomyocytes as context, while treating the latter as low rather than negative (HPA tissue IHC). Titrate the IHC validated antibody under fixed EDTA pH 8.0 retrieval conditions, and document the antibody SKU because the supplied caption names A16132-1 (datasheet A06348-1; selected tissue IHC caption; standard IHC practice).
How should I score PSPC1 staining across samples with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Score viable nuclei within a defined cell population rather than averaging whole sections with different proportions of cell types (HPA tissue IHC; standard IHC practice). Report the percentage of positive nuclei and, if intensity is reproducible, an H score from 0–300 using 0–3 intensity categories (standard IHC practice). Normalise each percentage to the number of evaluable nuclei in that same cell population and report the denominator (standard IHC practice). Keep retrieval, DAB development, counterstain and scoring thresholds consistent across the comparison, with blinded review where practical (standard IHC practice). Record tissue and cell type because HPA reports high glandular staining in appendix and low cardiomyocyte staining in heart muscle (HPA tissue IHC).
When should an apparent PSPC1 positive IHC result be questioned?
A convincing result should show staining in viable nuclei with a plausible cell type distribution; broad nuclear expression is supported in tissue IHC (HPA tissue IHC; standard IHC practice). Examine isolated cytoplasmic colour cautiously, while recognising that UniProt also lists cytoplasmic localisation for PSPC1 (UniProt Q8WXF1; standard IHC practice). Reject edge restricted staining, necrotic deposits or colour reproduced in the no primary control as evidence of specific nuclear signal (standard IHC practice). Check the peroxidase block when DAB appears in enzyme rich areas without matching nuclear morphology (standard IHC practice). Finally, the supplied paraffin tissue caption identifies antibody A16132-1, so its image and 2 μg/ml concentration do not establish performance for page SKU A06348-1 (selected tissue IHC caption).
Boster reagents
Best PSPC1 / Paraspeckle component 1 IHC Antibodies
Catalog antibody A06348-1 lists human paraffin-section IHC and human IF/ICC, with image captions for glioblastoma, ovarian serous adenocarcinoma, and U2OS cells (catalog applications; image captions).
Real IHC data
Anti-PSPC1 Antibody ®
Cat # A06348-1
A06348-1 lists IHC for human paraffin sections, with captions showing glioblastoma and ovarian serous adenocarcinoma tissue (catalog dilution data; IHC image captions). The same SKU lists human IF/ICC and has an IF caption for U2OS cells (catalog dilution data; IF image caption).
Which to pick: For tissue IHC, A06348-1 lists a starting concentration of 2–5 μg/ml; its paraffin-section captions describe EDTA retrieval at pH 8.0, although they name a different antibody code and do not report the fixative (catalog dilution data; IHC image captions). For IF/ICC, A06348-1 lists 5 μg/ml and has a U2OS IF caption (catalog dilution data; IF image caption). For cross-species work, A06348-1 lists human, mouse, and rat reactivity, while its IHC and IF/ICC dilution entries specify human samples (catalog reactivity; catalog dilution data).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8WXF1 (PSPC1_HUMAN, Paraspeckle component 1).
- Human Protein Atlas. PSPC1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PSPC1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli fibrillar center..
- Human Protein Atlas. PSPC1 antibody validation summary (1 antibodies).
- Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles. Molecular biology of the cell 2015 — PMC4395133.
- The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer. Cancer management and research 2021 — PMC8053714.
- PSPC1 is a potential prognostic marker for hormone-dependent breast cancer patients and modulates RNA processing of ESR1 and SCFD2. Scientific reports 2022 — PMC9184599.
- PSPC1-interchanged interactions with PTK6 and β-catenin synergize oncogenic subcellular translocations and tumor progression. Nature communications 2019 — PMC6914800.
- PubMed PMID:11790299 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15057823 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PSPC1 (Paraspeckle component 1). Expected localisation Predominantly nuclear in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06348-1). Validated IHC antibody: A06348-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative None in HPA (detected in all 44 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A06348-1); 2. Fixation: Image fixative and duration unreported (datasheet A06348-1); 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 A06348-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06348-1); 8. Primary antibody: Rabbit anti-PSPC1, 2-5 μg/ml (datasheet A06348-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06348-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Ovarian stromal cells show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: No intensity regulator specified (UniProt). Evidence: UniProt Q8WXF1, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06348-1, PMC4395133, PMC8053714. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PSPC1/immunohistochemistry-ihc-guide.