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PCBP1 / Poly(rC)-binding protein 1 · IHC design guide
Design Immunohistochemistry for PCBP1
Plan chromogenic PCBP1 staining in paraffin sections with the catalog antibody at 2–5 µg/ml (datasheet A02636-1). Use HPA tissue staining to assess the expected cytoplasmic pattern and tissue-dependent nuclear signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PCBP1 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; nuclear in some tissues (HPA tissue IHC) | |
| Staining pattern | Glandular, neuronal and adipocyte cytoplasm; some nuclei (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02636-1) | |
| 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 | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium concordance with tissue RNA (HPA tissue IHC) | |
| Regulation | No specific regulator annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–356 chain (UniProt) |
Section 1
Recommended PCBP1 IHC & IF Protocols
Compare the catalog antibody’s IHC protocol (datasheet A02636-1) with one published paraffin-section PCBP1 protocol (PMC9161940).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A02636-1) |
| Fixation | Image fixative and duration unreported (datasheet A02636-1); 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 A02636-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02636-1) |
| Primary antibody | Rabbit anti-PCBP1, 2-5μg/ml (datasheet A02636-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02636-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02636-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PCBP1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression with additional nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A02636-1); the published protocol does not specify retrieval (PMC9161940).
Section 2
What Is the Expected PCBP1 Staining Pattern?
In paraffin-section IHC, expect PCBP1 staining throughout the cytoplasm, with additional nuclear staining in some tissues (HPA: tissue IHC profile; UniProt Q15365: nucleus and cytoplasm). High staining is documented in selected glandular, neuronal, Purkinje and adipocyte populations (HPA: tissue IHC). The HPA IHC profile is Approved, with medium consistency between staining and RNA data (HPA: reliability). PCBP1 has no transmembrane segment (UniProt Q15365: topology).
| Cytoplasmic staining in adrenal, appendix, colon or duodenal glandular cells; nuclei may also stain. | This fits the reported ubiquitous cytoplasmic pattern with nuclear expression in several tissues (HPA: tissue IHC profile). Those glandular populations are rated High, but the record does not specify which individual nuclei should be positive (HPA: High in glandular cells). |
| Cytoplasmic staining in cortical or caudate neurons, cerebellar Purkinje cells, or breast adipocytes. | These are documented High-staining cell populations (HPA: tissue IHC). Interpret intensity against neighboring morphology and a matched control; High is an HPA category, not a calibrated chromogen threshold (HPA: tissue IHC levels). |
| Predominant sharp plasma-membrane outlines with little intracellular signal. | That compartment is discordant with nuclear and cytoplasmic localization and the absence of a transmembrane segment (UniProt Q15365: subcellular location and topology; HPA: tissue IHC profile). Review morphology, detection background and antibody specificity before scoring it as PCBP1. |
| Strong staining confined to an unexpected cell population while documented High-staining cells remain blank. | Cross-reactivity or endogenous detection activity is possible; check controls and the stained cell morphology (standard IHC practice). Other cell types are not automatically negative because HPA describes ubiquitous expression and lists no negative tissue controls (HPA: tissue IHC profile). |
| Diffuse haze, precipitate or widespread color without recognizable cellular compartments. | This cannot establish PCBP1 localization (standard IHC interpretation). Compare a no-primary control and inspect the chromogen distribution; the HPA reference pattern is intracellular, predominantly cytoplasmic, with nuclear expression in several tissues (HPA: tissue IHC profile). |
💡Expected PCBP1 appearanceCall a section positive when recognizable cells show cytoplasmic staining, with possible nuclear staining, including High signal in the listed HPA cell populations; isolated membrane outlines or diffuse cell-free color do not match the reference pattern (HPA: tissue IHC profile and High cell populations; UniProt Q15365: topology).
| Reference tissue and cell population | HPA rates glandular cells in adrenal gland, appendix, colon and duodenum High; it also rates breast adipocytes, caudate and cortical neurons, and cerebellar Purkinje cells High (HPA: tissue IHC). These are useful positive references, not an exhaustive list of expressing cells (HPA: ubiquitous profile). |
| Compartment and target structure | PCBP1 is annotated in nucleus and cytoplasm and may shuttle between them (UniProt Q15365: subcellular location). Its lack of a transmembrane segment gives no basis for a membrane-only call (UniProt Q15365: topology). Nuclear intensity can vary across tissues (HPA: tissue IHC profile). |
| Strength of the IHC reference | The tissue IHC profile is Approved, with medium consistency between antibody staining and RNA expression; CAB037113 is IHC Approved (HPA: tissue IHC reliability; HPA: antibody validation). Treat discrepant fields as cases for control review, rather than assuming every cell must match one intensity. |
| IF/ICC Q: Should paraffin IHC show nuclear speckles? | A: Nuclear speckles are the main supported ICC-IF location; nucleoplasm and cytosol are also supported, and cytoplasmic bodies are approved (HPA: subcellular ICC-IF). The tissue IHC description supports cytoplasmic staining with nuclear staining in several tissues, without requiring resolvable speckles in chromogenic sections (HPA: tissue IHC profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in adrenal or colon glandular cells. | These cells are rated High (HPA: tissue IHC); a blank section raises a workflow or reagent question, although HPA does not identify its cause. | Verify tissue identity and run a known-positive section; review the catalog antibody's IHC-P instructions, retrieval, primary dilution and detection controls (standard IHC practice). |
| Only nuclei stain across a section. | Nuclear PCBP1 is plausible (UniProt Q15365: subcellular location), but exclusive nuclear staining conflicts with the HPA ubiquitous cytoplasmic profile (HPA: tissue IHC). | Check cytoplasmic visibility and counterstain, compare a documented High cell population, and assess a matched control before assigning localization (standard IHC practice). |
| Crisp membrane staining dominates. | A membrane-only pattern conflicts with the annotated nucleus/cytoplasm location and absent transmembrane segment (UniProt Q15365: subcellular location and topology). | Inspect section edges and nonspecific chromogen, then compare the same cell type in a known-positive control (standard IHC practice). |
| An unexpected cell type stains intensely. | Cross-reactivity or endogenous detection activity may explain isolated staining (standard IHC practice); HPA's ubiquitous profile alone cannot classify that cell type as negative (HPA: tissue IHC). | Use a no-primary control, inspect cell identity and compare the documented High populations before attributing the signal to PCBP1 (standard IHC practice). |
| The whole section has diffuse brown background. | Nonspecific antibody binding, endogenous enzyme activity or chromogen deposits can obscure intracellular signal (standard chromogenic IHC practice). | Check a no-primary control, blocking and wash steps, detection reagent handling, and whether identifiable cytoplasm remains above background (standard IHC practice). |
| A proposed negative-control tissue also stains. | The supplied HPA record lists no negative tissues and describes ubiquitous cytoplasmic expression (HPA: tissue IHC profile). Tissue staining alone cannot validate that proposed negative control. | Use assay controls appropriate to the detection system and assess compartment and cell morphology against the documented positive pattern (standard IHC practice; HPA: tissue IHC profile). |
Experiment evidence
IHC & IF images showing PCBP1 staining
A02636-1 · IHCIHC analysis of PCBP1 using anti-PCBP1 antibody (A02636-1). PCBP1 was detected in a paraffin-embedded section of human breast cancer tissue.
A02636-1 · IHCIHC analysis of PCBP1 using anti-PCBP1 antibody (A02636-1). PCBP1 was detected in a paraffin-embedded section of human prostate cancer tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169564-PCBP1/tissue/cerebral+cortex#imid_9229192
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169564-PCBP1/tissue/colon#imid_9229118
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169564-PCBP1/tissue/liver#imid_9229119
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000169564-PCBP1/tissue/kidney#imid_9229120
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043803; green = PCBP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043803; green = PCBP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PCBP1 IHC & IF
🧪Run breast first and score adipocytes, which show High PCBP1 staining (HPA: breast adipocytes, High). HPA detects PCBP1 in all 45 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and expect cells without specific signal on the positive slide to show counterstain without DAB deposition, while treating them only as background references (HPA: no negative tissues; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: PCBP1 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 PCBP1 in A-431, U-251MG, U2OS, CACO-2, MCF-7, NIH 3T3, with annotated localisation: Nuclear speckles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a rabbit IgG isotype matched to the primary antibody’s clonality; use PCBP1 knockout material as a biological negative if available (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and inspect breast sections for nonspecific DAB background (standard chromogenic IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A02636-1 breast cancer paraffin-section caption does not state the fixative (caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 is documented for that IHC example, but a requirement for retrieval has not been established; whether frozen sections or IF are easier is unreported (caption: EDTA retrieval; HPA: ICC-IF images). In paraffin breast sections, adipocytes can appear as empty lipid spaces with thin cytoplasmic rims, so assess staining at the cell rim carefully (HPA: breast adipocytes, High; standard histology practice).
HPA tissue IHC evidence for PCBP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PCBP1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PCBP1 IHC Tips
Troubleshoot PCBP1 staining in paraffin sections by checking retrieval, compartment patterns, controls and scoring; use the IF guidance when planning a separate fluorescence assay.
What retrieval should I start with for PCBP1 in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A02636-1). The selected paraffin-section image used this retrieval before overnight primary-antibody incubation at 4°C (datasheet A02636-1). If staining is weak, vary heating time in small steps while keeping buffer and detection conditions fixed; excessive heating can damage section morphology (standard IHC practice). If EDTA remains ineffective, test an alternative retrieval buffer on adjacent sections as a fallback and compare signal with tissue integrity (standard IHC practice). Evaluate cytoplasmic and nuclear staining separately because both are plausible PCBP1 locations (UniProt Q15365; HPA tissue IHC).
How should I troubleshoot possible fixation effects on PCBP1 staining?
PCBP1-specific fixation sensitivity is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (datasheet A02636-1). Record each specimen’s fixative and fixation duration before comparing staining across cases, because fixation can alter antigen accessibility in IHC (standard IHC practice). When signal varies, stain matched sections in one run using the same pH 8.0 EDTA retrieval and primary-antibody conditions (datasheet A02636-1; standard IHC practice). Compare morphology and staining in cytoplasm and nuclei rather than assigning every difference to fixation (UniProt Q15365; standard IHC practice). Do not infer fixation tolerance from PCBP1’s tissue distribution or annotated modifications (HPA tissue IHC; UniProt Q15365).
Should PCBP1 stain nuclei, cytoplasm, or both in chromogenic IHC?
Expect cytoplasmic staining, with nuclear staining possible in some tissues: that is the reported tissue-IHC pattern (HPA tissue IHC). PCBP1 is annotated in both nucleus and cytoplasm and may shuttle between them (UniProt Q15365). Score the two compartments separately, using a counterstain to identify nuclei and tissue morphology to define the cells assessed (standard IHC practice). Do not require visible nuclear speckles in a DAB section; that finer pattern comes from subcellular ICC/IF evidence (HPA subcellular; standard IHC practice). If staining appears exclusively in an unexpected compartment, repeat with omission and independent specificity controls before interpreting a biological redistribution (standard IHC practice).
How can epitope uncertainty affect PCBP1 IHC interpretation?
The supplied PCBP1 record annotates a single chain of 356 residues and 0 isoforms; it does not establish this antibody’s precise epitope (UniProt Q15365). PCBP1 contains three KH domains at residues 13–75, 97–162 and 279–343 (UniProt Q15365). Annotated phosphoserines include residues 173, 189, 190 and 246, but their effect on this IHC antibody is untested here (UniProt Q15365). If staining changes between specimens, first standardise retrieval and staining conditions, then check antibody epitope information before proposing epitope masking (standard IHC practice). Use an independent antibody or orthogonal assay to assess specificity when the compartment pattern remains unexplained (standard IHC practice).
How should I adapt PCBP1 localisation checks for multiplex IF?
Use this as planning guidance for the separate IF/ICC assay; the selected antibody caption documents paraffin-section chromogenic IHC, not IF performance (datasheet A02636-1). Multiplex PCBP1 with a validated marker for the expected cell type, and choose fluorophores whose signals can be distinguished from tissue autofluorescence and one another (standard IF practice). Include single-stain controls to assess spectral bleed-through before assigning PCBP1 to a marked cell (standard IF practice). Because PCBP1 is annotated in nucleus and cytoplasm and lacks a transmembrane segment, optimise permeabilisation to reveal intracellular epitopes without disrupting morphology (UniProt Q15365; standard IF practice). Compare nuclear and cytoplasmic signal while recognising that nuclear speckles are supported by subcellular ICC/IF evidence (HPA subcellular).
What should I check when PCBP1 DAB staining is diffuse?
First inspect a section processed without primary antibody to distinguish detection-system background from antibody-dependent staining (standard IHC practice). The selected workflow used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, a biotinylated secondary and DAB development (datasheet A02636-1). Apply an appropriate peroxidase block and control DAB development time, since endogenous enzyme activity and overdevelopment can create misleading colour (standard IHC practice). If background persists, titrate the primary antibody around the documented concentration and review blocking, washing and section drying (datasheet A02636-1; standard IHC practice). Retain distinct cytoplasmic or nuclear signal only when controls and morphology support it (HPA tissue IHC; standard IHC practice).
How should I quantify PCBP1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then report nuclear and cytoplasmic staining separately because both patterns are plausible (UniProt Q15365; HPA tissue IHC; standard IHC practice). For intensity-based assessment, use a compartment-specific H-score calculated as the sum of each intensity category multiplied by its percentage of cells (standard IHC practice). Alternatively, report the percentage of positive cells or positive-cell density per mm² of viable tissue, with a prespecified positivity threshold (standard IHC practice). Normalise counts to the number of eligible cells or measured viable area, and keep imaging and DAB development settings consistent (standard IHC practice). Record cell type, since HPA reports high staining in selected glandular, adipocyte and neuronal populations (HPA tissue IHC).
How can I distinguish convincing PCBP1 staining from artefact?
Treat cytoplasmic staining, with possible nuclear staining, as the expected compartment framework for PCBP1 tissue IHC (HPA tissue IHC; UniProt Q15365). Judge the cell population against the tissue context: HPA reports high signal in adrenal and colon glandular cells, breast adipocytes and cortical neuronal cells (HPA tissue IHC). Be cautious with signal confined to section edges, damaged or necrotic areas, or the no-primary control, which can indicate processing or detection artefact (standard IHC practice). Check peroxidase blocking when brown deposit appears without a convincing cell pattern (standard IHC practice). HPA rates its tissue staining Approved with medium consistency against RNA, so confirm surprising patterns independently (HPA tissue IHC; standard IHC practice).
Boster reagents
Best PCBP1 / Poly(rC)-binding protein 1 IHC Antibodies
A02636-1 has real PCBP1 IHC images from human paraffin sections of breast, prostate and rectal cancer and tonsil (catalog IHC image captions). No IF images are supplied (catalog IF image alts).
Real IHC data
Anti-PCBP1 Antibody ®
Cat # A02636-1
A02636-1 is listed for IHC and shows staining in human paraffin sections of breast, prostate and rectal cancer and tonsil (catalog applications; IHC image captions). Its listed reactivity is human, mouse and rat, while the supplied IHC images show human tissue only (catalog reactivity; IHC image captions).
Which to pick: Choose A02636-1 for human paraffin-section IHC; its own captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml, but do not report the fixative (catalog IHC image captions). No SKU can be recommended here for IF/ICC because A02636-1 has no listed IF application or IF image (catalog applications; IF image alts). For cross-species work, A02636-1 lists mouse and rat reactivity, but its supplied IHC examples cover human tissue only (catalog reactivity; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15365 (PCBP1_HUMAN, Poly(rC)-binding protein 1).
- Human Protein Atlas. PCBP1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PCBP1 subcellular location (ICC-IF): Mainly localized to the nuclear speckles. In addition localized to the nucleoplasm, cytosol and cytoplasmic bodies..
- Human Protein Atlas. PCBP1 antibody validation summary (3 antibodies).
- Generation of PCBP1-deficient pigs using CRISPR/Cas9-mediated gene editing. iScience 2022 — PMC9579030.
- Mycobacterium tuberculosis-induced PCBP1 degradation drives macrophage ferroptosis to promote infection: a lung-macrophage-targeted RNAa nanotherapy. Journal of nanobiotechnology 2026 — PMC13377778.
- PolyC-RNA-binding protein 1 (PCBP1) enhances tropomyosin 3 (TPM3) mRNA stability to promote the progression of esophageal squamous cell carcinoma. Bioengineered 2022 — PMC9161940.
- Poly C Binding Protein 1 Regulates p62/SQSTM1 mRNA Stability and Autophagic Degradation to Repress Tumor Progression. Frontiers in genetics 2020 — PMC7457068.
- PubMed PMID:7607214 — UniProt-cited evidence.
- PubMed PMID:7556077 — UniProt-cited evidence.
- PubMed PMID:8152927 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PCBP1 (Poly(rC)-binding protein 1). Expected localisation Cytoplasmic; nuclear in some tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02636-1). Validated IHC antibody: A02636-1 at 2-5μg/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 breast cancer tissue; fixative not specified (datasheet A02636-1); 2. Fixation: Image fixative and duration unreported (datasheet A02636-1); 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 A02636-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02636-1); 8. Primary antibody: Rabbit anti-PCBP1, 2-5μg/ml (datasheet A02636-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02636-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining has medium concordance with tissue RNA (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–356 chain (UniProt); Regulation: No specific regulator annotated (UniProt). Evidence: UniProt Q15365, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nuclear speckles. In addition localized to…, Datasheet A02636-1, PMC9161940. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PCBP1/immunohistochemistry-ihc-guide.