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PABPC4 / Polyadenylate-binding protein 4 · IHC design guide
Design Immunohistochemistry for PABPC4
Plan paraffin-section PABPC4 staining around cytoplasmic signal in most tissues, including immune-cell subsets (HPA tissue IHC). The catalog antibody has a documented chromogenic IHC-P application; compare staining by cell type with the HPA tissue profile (datasheet A07960-2; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PABPC4 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 | Cytoplasmic in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues, including immune-cell subsets (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07960-2) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | No source-confirmed negative tissue/cell row; 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 | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | T-cell activation rapidly raises mRNA (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended PABPC4 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A07960-2) is accompanied by published PABPC4 staining methods for prostate cancer TMAs (PMC12336731) and colorectal tissues (PMC13496150).
| Sample | Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A07960-2) |
| Fixation | Image fixative and duration unreported (datasheet A07960-2); 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 A07960-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07960-2) |
| Primary antibody | Rabbit anti-PABPC4, 2-5 μg/ml (datasheet A07960-2) |
| Primary incubation | Overnight at 4 °C (datasheet A07960-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07960-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PABPC4-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, including subsets of immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet: A07960-2); citrate retrieval is a published alternative for colorectal sections (PMC13496150).
Section 2
What Is the Expected PABPC4 Staining Pattern?
PABPC4 is a cytoplasmic protein with no transmembrane segment (UniProt Q13310 topology). In paraffin sections, expect cytoplasmic staining in many tissues, including subsets of immune cells (HPA: tissue IHC profile). Strong examples include cerebellar Purkinje cells, pancreatic exocrine glandular cells, and placental cytotrophoblasts (HPA: High). HPA rates the tissue IHC pattern Supported, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Cytoplasmic stain in Purkinje cells, pancreatic exocrine cells, or placental cytotrophoblasts. | This matches reported High staining in those cell types (HPA: tissue IHC). Compare each cell population with nearby tissue before scoring: PABPC4 has a broad cytoplasmic profile, so staining outside these examples is not automatically unexpected (HPA: tissue IHC profile). |
| A strong nuclear-only, membrane-rim, or extracellular pattern dominates the section. | That compartment conflicts with the expected cytoplasmic localization (UniProt Q13310 subcellular location; HPA: tissue IHC profile). Treat it as suspect and review morphology, detection controls, and staining conditions (general IHC practice). UniProt reports cytoplasmic mRNP granules, but that does not establish a required punctate IHC pattern (UniProt Q13310 subcellular location). |
| The dominant stain is in cells other than the expected population for the chosen tissue. | Compare the stained cells with the tissue-specific HPA entry before calling the result positive (HPA: tissue IHC). An unexpected distribution can reflect antibody cross-reactivity or endogenous detection activity (general IHC practice); it cannot be diagnosed from location alone, especially given the broad reported expression (HPA: tissue IHC profile). |
| Diffuse color covers tissue and blank areas, obscuring cell boundaries. | A field-wide haze prevents a reliable cytoplasmic call (general IHC practice). Check background on a no-primary control, then review blocking, washes, and chromogen development (general IHC practice). HPA reports staining by cell population; it does not identify a PABPC4-specific cause of diffuse background (HPA: tissue IHC). |
| No discernible stain appears in a section selected for reported High expression. | Confirm that the section contains the relevant cells: HPA reports High staining in pancreatic exocrine glandular cells and cerebellar Purkinje cells (HPA: tissue IHC). Then check the staining run and tissue integrity (general IHC practice). A blank result alone does not establish PABPC4 absence (HPA: Supported, medium RNA–protein consistency). |
💡Expected PABPC4 appearanceCall a result positive when the relevant cells show discernible cytoplasmic chromogen, potentially strong in HPA High populations; dominant nuclear-only, membrane-rim, or field-wide stain is suspect (UniProt Q13310 subcellular location; HPA: tissue IHC levels; general IHC practice).
| Tissue and cell population | HPA reports High staining in Purkinje cells, ovarian follicle cells, pancreatic exocrine glandular cells, placental cytotrophoblasts, and spermatogonia (HPA: tissue IHC). It lists Low staining in hepatocytes and several epithelial or glandular populations (HPA: tissue IHC); compare like cells when judging intensity. |
| Expected compartment and topology | Cytoplasm is the primary location, with cytoplasmic mRNP granules also described (UniProt Q13310 subcellular location). The protein has no transmembrane segment or signal peptide (UniProt Q13310 topology and processing), so a dominant membrane-rim or secreted-looking pattern needs scrutiny. |
| Evidence strength | The tissue profile is Supported, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). HPA lists IHC as Supported for HPA027301 and HPA056496; their Enhanced status applies to ICC, not IHC (HPA: antibody validation). |
| Isoforms and antibody recognition | UniProt lists three PABPC4 isoforms (UniProt Q13310 isoforms). The supplied record gives no epitope location or isoform-specific IHC result, so it cannot establish which isoforms an antibody detects in sections (UniProt Q13310 isoforms; HPA: antibody validation). |
| IF/ICC cross-check: what location is expected? | HPA reports enhanced cytosol localization in ICC-IF and Enhanced ICC validation for HPA027301 and HPA056496 (HPA: subcellular; HPA: antibody validation). Use that as compartment context; it does not supply an IHC-P retrieval condition or establish equal performance between applications. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported High tissue is blank. | The relevant cells may be absent from the section, or the staining run may have failed (HPA: tissue IHC; general IHC practice). | Check for the specified cells, then review tissue integrity, the run control, detection reagents, and retrieval against the IHC-validated antibody's instructions (general IHC practice). No PABPC4-specific retrieval setting is supplied. |
| Signal is weak in a reported Low population. | HPA lists Low staining in hepatocytes, lung type II alveolar cells, and several glandular or epithelial populations (HPA: tissue IHC). | Compare the same cell type across sections and include a reported High population when available (HPA: tissue IHC; general IHC practice). Do not turn a weak Low-population result into an assay failure without a run control. |
| Nuclear-only or membrane-rim stain dominates. | The distribution conflicts with cytoplasmic localization and lack of a transmembrane segment (UniProt Q13310 subcellular location and topology). | Review morphology and a no-primary control; check antibody dilution and detection conditions under the applicable IHC protocol (general IHC practice). Do not score the discordant compartment as the expected PABPC4 pattern. |
| Many unrelated structures carry similar diffuse chromogen. | Nonspecific binding, endogenous detection activity, or excess chromogen development are possible general IHC causes; this appearance is not a PABPC4-specific finding (general IHC practice). | Compare a no-primary control, verify appropriate blocking, and review wash and development steps (general IHC practice). Interpret cell-specific cytoplasmic staining only after background is controlled. |
| An unexpected cell type stains more strongly than the reported High population. | Cross-reactivity or endogenous detection activity is possible, but HPA also reports cytoplasmic expression in most tissues (general IHC practice; HPA: tissue IHC profile). | Identify the stained cells histologically, compare their HPA tissue entry, and inspect detection controls (HPA: tissue IHC; general IHC practice). Seek independent antibody support before assigning an unexpected pattern to PABPC4 (general IHC practice). |
| Results vary between sections after retrieval changes. | Retrieval is a general IHC variable; the supplied sources do not report PABPC4-specific fixation sensitivity or an optimal retrieval condition (general IHC practice; HPA: tissue IHC; UniProt Q13310). | Hold tissue and detection conditions consistent while following the IHC-validated antibody's protocol, and compare run controls (general IHC practice). Report the observed conditions without attributing the difference to a proven PABPC4 fixation effect. |
Experiment evidence
IHC & IF images showing PABPC4 staining
A07960-2 · IHCIHC analysis of APP-1/PABPC4 using anti-APP-1/PABPC4 antibody (A07960-2). APP-1/PABPC4 was detected in a paraffin-embedded section of human
A07960-2 · IHCIHC analysis of APP-1/PABPC4 using anti-APP-1/PABPC4 antibody (A07960-2). APP-1/PABPC4 was detected in a paraffin-embedded section of human
A07960-2 · IFIF analysis of APP-1/PABPC4 using anti-APP-1/PABPC4 antibody (A07960-2). APP-1/PABPC4 was detected in an immunocytochemical section of SiHa
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000090621-PABPC4/tissue/cerebral+cortex#imid_7231584
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000090621-PABPC4/tissue/colon#imid_7231526
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000090621-PABPC4/tissue/liver#imid_7231511
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000090621-PABPC4/tissue/kidney#imid_7231528
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027301; green = PABPC4, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027301; green = PABPC4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PABPC4 IHC & IF
🧪Run cerebellum first; Purkinje cells should show cytoplasmic staining because HPA rates them High for PABPC4 (HPA: High in Purkinje cells; UniProt: cytoplasmic localisation). No source-confirmed negative tissue or internal negative cell type is listed among the supplied HPA candidates; use no-primary and isotype controls to establish background, and treat any unstained neighboring cells as background comparators rather than validated PABPC4-negative cells (HPA: no negative rows supplied).
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: No source-confirmed negative tissue/cell row among selected HPA candidates. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PABPC4 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a matched nonimmune rabbit IgG control, alongside a PABPC4 knockout biological negative where available (selected-SKU caption: rabbit primary antibody; standard IHC control practice). Quench endogenous peroxidase before HRP/DAB detection and assess pigment in the cerebellar section when judging signal (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative unreported). The caption uses heat-mediated retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; the supplied evidence does not establish whether frozen sections or IF are easier (selected-SKU caption: EDTA retrieval; HPA: ICC-IF images in A-431, U-251MG and U2OS). PABPC4 is cytoplasmic, so IF/ICC assessment should allow access to intracellular antigen through permeabilisation; cerebellar neuronal pigment or autofluorescence may complicate interpretation (UniProt: cytoplasmic localisation; standard IF practice).
HPA tissue IHC evidence for PABPC4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Cytotrophoblasts | High | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No source-confirmed negative tissue/cell row among selected HPA candidates. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PABPC4 IHC Tips
Use cytoplasmic staining, tissue morphology and matched controls to evaluate PABPC4 in paraffin section IHC (UniProt Q13310 localisation; standard IHC practice).
How should I optimize antigen retrieval when PABPC4 staining is weak?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A07960-2). The selected image used this retrieval with 2 μg/ml primary antibody overnight at 4°C in human cervical cancer tissue; its caption does not report a fixative (caption A07960-2). If staining is weak, compare heating durations on serial sections while holding antibody concentration, incubation and detection constant; test another retrieval buffer only as a fallback (standard IHC practice). Select the condition that preserves morphology and gives cellular cytoplasmic signal, the expected compartment for PABPC4 (UniProt Q13310 localisation).
Can I use the published staining pattern to choose a fixation time?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative or fixation time (caption A07960-2). Record each specimen’s fixative, time in fixative and processing schedule before comparing staining, since these variables can affect IHC results (standard IHC practice). On matched sections, keep EDTA retrieval at pH 8.0 and hold antibody incubation and chromogenic detection constant (datasheet A07960-2; standard IHC practice). Evaluate preserved architecture, cytoplasmic signal and a negative reagent control together; neither a tissue staining pattern nor PABPC4’s topology establishes how fixation affects this antigen (UniProt Q13310 localisation and topology; standard IHC practice).
Should punctate or nuclear staining count as PABPC4 signal?
Expect predominantly cytoplasmic staining in intact cells: PABPC4 is cytoplasmic and can localize to cytoplasmic mRNP granules (UniProt Q13310 localisation). The tissue profile also describes cytoplasmic expression across most tissues, while its staining reliability is Supported with medium RNA concordance (HPA tissue IHC). Score a reproducible cytoplasmic punctate pattern separately from diffuse cytoplasmic staining, using the same retrieval and DAB development for comparison (UniProt Q13310 localisation; standard IHC practice). Treat isolated nuclear staining cautiously and check serial sections, morphology and a negative reagent control before assigning it to PABPC4 (UniProt Q13310 localisation; standard IHC practice).
Could isoforms or epitope position change the IHC result?
PABPC4 has 3 recorded isoforms and four RNA recognition motifs, followed by a PABC domain at residues 551–628 (UniProt Q13310 isoforms and domains). The supplied antibody caption does not identify its epitope, so it does not establish which isoforms the catalog antibody detects (caption A07960-2). Request the immunogen or epitope sequence and map it against the recorded isoforms before interpreting an apparent absence of staining (UniProt Q13310 isoforms; standard IHC practice). Several modified residues are recorded, but their presence alone cannot establish altered epitope accessibility in paraffin sections; resolve that experimentally with appropriate specificity controls (UniProt Q13310 modified residues; standard IHC practice).
How should I plan IF validation alongside the chromogenic IHC result?
Treat IF/ICC as a separate validation, while using the IHC result to define the expected cellular compartment: PABPC4 is cytoplasmic, with an enhanced cytosol location in the subcellular profile (UniProt Q13310 localisation; HPA subcellular). For multiplex IF, pair PABPC4 with a validated marker for the expected cell type, such as exocrine glandular cells when examining pancreas (HPA tissue IHC; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence and keep channels spectrally separated (standard IF practice). Because the target is intracellular and has no transmembrane segment, validate permeabilisation for access to its cytoplasmic epitope, with matched single-channel and secondary-only controls (UniProt Q13310 topology; standard IF practice).
How can I separate diffuse DAB background from PABPC4 staining?
The selected paraffin-section image used 10% goat serum blocking, a 2 μg/ml primary antibody overnight at 4°C, and DAB development (caption A07960-2). Begin troubleshooting with a no-primary control, a peroxidase block and matched DAB development times to locate reagent or endogenous enzyme background (standard chromogenic IHC practice). Titrate the primary and shorten chromogen development on adjacent sections if diffuse colour obscures cell boundaries, while retaining the same retrieval condition (datasheet A07960-2; standard IHC practice). Count only staining that resolves within intact cytoplasm and exceeds the negative control, consistent with PABPC4’s reported location (UniProt Q13310 localisation; standard IHC practice).
What should I measure when PABPC4 staining varies across cells? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population and tissue region before scoring, then record the percentage of cytoplasm-positive cells and an intensity-weighted H-score under one fixed scoring rule (standard IHC practice). For dispersed positive cells, report their density per mm² of viable tissue and normalize positive counts to the number of eligible cells when cell density differs (standard IHC practice). Keep retrieval, exposure to primary antibody and DAB development consistent across sections, and set positivity against a matched negative reagent control (standard IHC practice). Report cell type separately because high staining is listed for pancreatic exocrine glandular cells and several other specified populations (HPA tissue IHC).
Which staining features support a true PABPC4-positive result?
A convincing result is reproducible cytoplasmic staining in intact cells that exceeds a matched negative reagent control, consistent with PABPC4’s reported cytoplasmic location (UniProt Q13310 localisation; standard IHC practice). Compare the stained cell type with the tissue profile: high staining is reported in pancreatic exocrine glandular cells, whereas hepatocytes are listed as low (HPA tissue IHC). Exclude tissue edges and necrotic regions from scoring if they show disproportionate colour or poor morphology, and inspect no-primary sections for endogenous peroxidase signal (standard chromogenic IHC practice). Interpret discordant nuclear-only or widespread uniform deposits cautiously, especially because the tissue staining profile has medium concordance with RNA expression (UniProt Q13310 localisation; HPA tissue IHC).
Boster reagents
Best PABPC4 / Polyadenylate-binding protein 4 IHC Antibodies
A07960-2 has IHC images from human paraffin sections and IF/ICC images from human tissue and SiHa cells (catalog image captions). Listed reactivity includes human, mouse, and rat (catalog).
Real IHC data
Anti-APP-1/PABPC4 Antibody ®
Cat # A07960-2
A07960-2 was demonstrated by IHC on paraffin sections of human cervical cancer and tonsil tissue (catalog IHC image captions). Its IF images show SiHa cells and paraffin sections of human chronic tonsillitis and ovary cancer tissue (catalog IF image captions).
Which to pick: For tissue IHC, choose A07960-2: its paraffin-section captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (catalog IHC image captions). For IF/ICC, the same SKU has human cell and tissue images at 5 μg/ml (catalog IF image captions). For mouse or rat samples, A07960-2 lists reactivity with both species, but the supplied IHC and IF images document human samples (catalog reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13310 (PABP4_HUMAN, Polyadenylate-binding protein 4).
- Human Protein Atlas. PABPC4 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PABPC4 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. PABPC4 antibody validation summary (2 antibodies).
- Elevated PABPC4 expression in human prostate cancer tissues predicts adverse clinical outcomes. Translational andrology and urology 2025 — PMC12336731.
- ENO1-related gene signature predicts prognosis and therapeutic response in diffuse large B-cell lymphoma. Frontiers in immunology 2025 — PMC12588913.
- Macrophage PABPC4-SPP1 Axis Orchestrates Immunosuppression in Colorectal Cancer. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13496150.
- PubMed PMID:8524242 — UniProt-cited evidence.
- PubMed PMID:9030741 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PABPC4 (Polyadenylate-binding protein 4). Expected localisation Cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07960-2). Validated IHC antibody: A07960-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative No source-confirmed negative tissue/cell row; use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A07960-2); 2. Fixation: Image fixative and duration unreported (datasheet A07960-2); 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 A07960-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07960-2); 8. Primary antibody: Rabbit anti-PABPC4, 2-5 μg/ml (datasheet A07960-2); 9. Primary incubation: Overnight at 4 °C (datasheet A07960-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unknown (UniProt); Regulation: T-cell activation rapidly raises mRNA (UniProt). Evidence: UniProt Q13310, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A07960-2, PMC12336731, PMC13496150. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PABPC4/immunohistochemistry-ihc-guide.