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PCBP1 / Poly(rC)-binding protein 1 · IHC design guide
Design Immunohistochemistry for PCBP1
Plan chromogenic IHC for PCBP1 in paraffin sections using the catalog antibody’s documented conditions (datasheet A02636-1). Use the broad cytoplasmic pattern and tissue dependent nuclear staining to guide interpretation (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 several tissues (HPA tissue IHC) | |
| Staining pattern | Broad cytoplasmic staining, with nuclear staining in some tissues (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 sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02636-1) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | No isoforms or processed chains are annotated (UniProt) |
Section 1
Recommended PCBP1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A02636-1) with the published ESCC tissue protocol below (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 pH 8.0 retrieval (datasheet A02636-1); the published ESCC protocol does not specify retrieval (PMC9161940).
Section 2
What Is the Expected PCBP1 Staining Pattern?
PCBP1 is a nuclear and cytoplasmic protein without a transmembrane segment (UniProt Q15365). In paraffin tissue sections, expect broadly distributed cytoplasmic staining, with nuclear staining in some tissues (HPA: ubiquitous cytoplasmic expression; additional nuclear expression in several tissues). HPA rates its tissue IHC evidence Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Cytoplasmic stain in glandular cells of colon, duodenum, appendix or adrenal gland. | This fits the tissue profile: HPA reports High staining in those cells and describes PCBP1 as ubiquitously cytoplasmic (HPA: tissue IHC). Nuclear stain may also be compatible, depending on the tissue (HPA: additional nuclear expression in several tissues). |
| Predominantly sharp membrane or extracellular stain, with little intracellular stain. | Treat this as a pattern mismatch and investigate artefact or nonspecific detection: PCBP1 is nuclear and cytoplasmic, with no transmembrane segment or signal peptide (UniProt Q15365). |
| Strong stain confined to an unexpected cell population while the expected cells remain unstained. | Consider cross-reactivity or endogenous detection activity (general IHC practice). HPA reports High glandular cell staining in colon, duodenum, appendix and adrenal gland; its supplied tissue profile lists no negative cell populations, so an unexpected positive alone does not prove cross-reactivity (HPA: tissue IHC). |
| Uniform color over tissue, stroma and empty areas, obscuring cell boundaries. | This is background rather than an interpretable PCBP1 compartment pattern (general IHC practice). Compare the no-primary control and inspect blocking, washes and detection chemistry (general IHC practice). |
| No intracellular signal in an otherwise intact known-positive section. | A section containing HPA High glandular cells should offer a positive reference (HPA: colon, duodenum, appendix and adrenal gland). First check the staining run and antibody conditions; HPA's supplied list contains no validated negative tissues for a reciprocal comparison (HPA: tissue IHC). |
💡Expected PCBP1 appearanceCall a section positive when identifiable cells show intracellular cytoplasmic stain, with possible nuclear stain, especially in HPA High glandular cells; isolated membrane or extracellular color is suspect (HPA: tissue IHC; UniProt Q15365).
| Which reference tissue and cells support interpretation? | HPA reports High staining in glandular cells of adrenal gland, appendix, colon and duodenum; it also lists High adipocytes in breast and neuronal cells in caudate and cerebral cortex (HPA: tissue IHC). These are observed positives, not a required intensity in every section. |
| How strong is the tissue IHC evidence? | The tissue profile is Approved, with medium agreement between antibody staining and RNA data (HPA: tissue IHC). CAB037113 is IHC Approved, while the supplied HPA records give no IHC status for HPA043803 or HPA068497 (HPA: antibody validation). |
| Does molecular location suggest membrane or secreted stain? | UniProt places PCBP1 in the nucleus and cytoplasm and reports no transmembrane segment, signal peptide or propeptide (UniProt Q15365). Interpret dominant surface or extracellular color cautiously; those features do not establish an antigen retrieval or fixation effect. |
| What should IF/ICC show? | HPA reports mainly nuclear speckles, with supported nucleoplasm and cytosol and approved cytoplasmic bodies in ICC-IF (HPA: subcellular). HPA043803 and HPA068497 have Enhanced ICC validation (HPA: antibody validation). Nuclear speckles are an IF observation, not a required chromogenic tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular cells are blank in a candidate positive section. | The run, detection reagents or primary antibody conditions may have failed (general IHC practice); HPA reports these cells as High in colon, duodenum, appendix and adrenal gland (HPA: tissue IHC). | Confirm tissue integrity and counterstain, then review the antibody's IHC-P instructions, detection reagents and run controls (general IHC practice). Do not infer PCBP1-specific fixation sensitivity from this result. |
| Color is widespread and cell boundaries are hard to distinguish. | Nonspecific primary or secondary binding, incomplete blocking, or inadequate washing can produce diffuse background (general IHC practice). | Compare a no-primary control; review blocking, wash steps and primary concentration for this chromogenic assay (general IHC practice). Score compartment staining only after background is controlled. |
| Brown signal appears without primary antibody. | Endogenous enzyme activity or nonspecific detection chemistry may contribute, depending on the detection system (general IHC practice). | Check the no-primary control and apply the detection system's appropriate endogenous-activity block; repeat with the same tissue and counterstain (general IHC practice). |
| Only membrane rims or extracellular deposits are conspicuous. | That distribution conflicts with PCBP1's nuclear and cytoplasmic annotation and lack of a transmembrane segment or signal peptide (UniProt Q15365). | Inspect no-primary and reagent controls, confirm the chromogen deposit lies within cells, and compare with HPA's intracellular tissue pattern (general IHC practice; HPA: tissue IHC). |
| Nuclear stain is present but discrete speckles are not visible in IHC. | Additional nuclear expression occurs in some tissues (HPA: tissue IHC); speckles are the main ICC-IF location (HPA: subcellular). Chromogenic tissue IHC need not resolve the same detail (general IHC practice). | Judge the tissue section by its intracellular staining and cell identity; use the HPA ICC-IF pattern only to answer the separate IF localisation question (HPA: tissue IHC; HPA: subcellular). |
| A different antibody gives a conflicting tissue pattern. | The supplied HPA validation differs by application: CAB037113 is IHC Approved; HPA043803 and HPA068497 have Enhanced ICC status but no stated IHC status (HPA: antibody validation). | Check each antibody's IHC-P validation and compare matched sections and controls before attributing the difference to biology (general IHC practice). |
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 tissue first and score adipocytes for PCBP1 staining (HPA: High in breast adipocytes). HPA detects PCBP1 in all 45 scored tissues, so there is no validated negative tissue or internal negative cell type; cells without signal should remain at background, but should not be designated biological negatives without validation (HPA: no negative rows).
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 slide, a rabbit IgG isotype control matched to the primary antibody where applicable, and a matched PCBP1 knockout specimen as a biological negative (selected-SKU caption: rabbit anti-PCBP1). For chromogenic detection, quench endogenous peroxidase and check for endogenous biotin background when using the caption’s biotinylated secondary and streptavidin–biotin detection system (selected-SKU caption: SABC with DAB).
⚠️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). Heat retrieval in EDTA at pH 8.0 produced the captioned IHC result, but retrieval dependency and whether frozen sections are easier are unreported (selected-SKU caption: EDTA pH 8.0). ICC-IF images support IF assessment of nuclear speckles, while breast adipose autofluorescence should be checked with an unstained section if IF is used (HPA subcellular: nuclear speckles supported; HPA: High in breast adipocytes).
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, controls and cell-level scoring before interpreting chromogenic signal.
What retrieval conditions should I start with for PCBP1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A02636-1). The selected paraffin-section image used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C (caption A02636-1). If staining is weak, compare retrieval heating and cooling across matched sections while holding antibody concentration and detection constant (standard IHC practice). Check whether signal appears in cytoplasm or nuclei, since both are reported PCBP1 compartments (UniProt Q15365; HPA tissue IHC). Include a section processed without primary antibody to distinguish retrieval-associated background from specific staining (standard IHC practice).
How should I troubleshoot weak staining when the tissue fixative is unknown?
The selected PCBP1 image documents a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (caption A02636-1). Record the available fixation history and compare sections with known, consistent processing before assigning weak signal to PCBP1 abundance (standard IHC practice). Keep retrieval at EDTA pH 8.0 and primary antibody at 2 μg/ml for the initial comparison (datasheet A02636-1; caption A02636-1). Examine intact central tissue as well as section edges, where processing artefacts can mislead visual assessment (standard IHC practice). Interpret remaining differences alongside staining controls and tissue morphology, without assuming a PCBP1-specific fixation effect (standard IHC practice).
Should PCBP1 staining be nuclear, cytoplasmic, or both?
Score nuclear and cytoplasmic staining separately because PCBP1 is reported in both compartments and may shuttle between them (UniProt Q15365). Tissue IHC shows widespread cytoplasmic expression with additional nuclear staining in several tissues (HPA tissue IHC). Cell imaging places PCBP1 mainly in nuclear speckles, with additional nucleoplasmic, cytosolic and cytoplasmic-body localisation (HPA subcellular). In chromogenic sections, compare signal with nuclear counterstain and cell boundaries before calling puncta or diffuse staining compartment-specific (standard IHC practice). Review matched controls when a case shows only extracellular signal or staining that follows damaged tissue rather than intact cells (standard IHC practice).
Could epitope accessibility or PCBP1 variants explain uneven staining?
No isoforms are annotated for the 356-amino-acid PCBP1 record, so do not assign different staining patterns to a named isoform (UniProt Q15365). PCBP1 contains 3 KH domains and several annotated modified residues, but the supplied antibody caption does not locate its epitope (UniProt Q15365; caption A02636-1). Compare staining after the stated EDTA pH 8.0 retrieval under otherwise matched conditions before proposing epitope masking (datasheet A02636-1; standard IHC practice). If another antibody with a documented, distinct epitope is available, concordant cellular staining can strengthen interpretation (standard IHC practice). Report persistent differences as unresolved antibody or processing effects unless epitope-specific evidence establishes a mechanism (standard IHC practice).
How can I check PCBP1 localisation by IF alongside chromogenic IHC?
Treat IF as a separate validation experiment: the selected antibody evidence documents paraffin-section chromogenic IHC, while subcellular imaging reports PCBP1 in nuclear speckles, nucleoplasm and cytosol (caption A02636-1; HPA subcellular). Multiplex PCBP1 with a validated marker for the expected cell type, and verify each channel individually before judging colocalisation (standard IF practice). Choose fluorophores and filters that separate specific signal from the specimen's autofluorescence, using unstained and single-label controls (standard IF practice). Because PCBP1 has no transmembrane segment and occupies nuclear and cytoplasmic compartments, use controlled permeabilisation to reach intracellular epitopes (UniProt Q15365; standard IF practice). Confirm the chosen IF fixation and antibody conditions experimentally (standard IF practice).
How do I distinguish PCBP1 signal from chromogenic background?
Run a no-primary control and inspect tissue edges, folds and damaged areas before assigning diffuse DAB colour to PCBP1 (standard IHC practice). In a peroxidase-based chromogenic workflow, block endogenous peroxidase and assess whether residual colour persists without primary antibody (standard IHC practice). The selected image used a biotinylated secondary, avidin-biotin detection and DAB, so include detection controls when troubleshooting that workflow (caption A02636-1). If broad staining overwhelms cell boundaries, compare blocking, washes and primary concentration on matched sections while keeping retrieval at EDTA pH 8.0 (datasheet A02636-1; standard IHC practice). Retain interpretable nuclear counterstain and morphology for compartment scoring (standard IHC practice).
What should I measure when comparing PCBP1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and eligible cell population before scoring, then measure nuclear and cytoplasmic PCBP1 separately (standard IHC practice; UniProt Q15365). For cell-based comparisons, report percentage of positive cells and an intensity-weighted H-score using the same thresholds across slides (standard IHC practice). Where cell density varies, report positive cells per mm² and normalise to the area of intact, evaluable tissue (standard IHC practice). Apply the same retrieval, staining batch, counterstain and image settings to comparison sections, including EDTA pH 8.0 retrieval (datasheet A02636-1; standard IHC practice). Exclude folds and necrotic regions by a prespecified rule (standard IHC practice).
When is a positive PCBP1 result biologically credible?
A credible result follows intact cell boundaries and shows cytoplasmic or nuclear staining, the compartments reported for PCBP1 (UniProt Q15365; HPA tissue IHC). Check cell identity before interpreting intensity: HPA reports high staining in adrenal glandular cells, breast adipocytes and several neuronal populations (HPA tissue IHC). Compare suspicious edge staining, necrotic regions and colour in the no-primary control with the candidate positive cells (standard IHC practice). Residual endogenous enzyme activity can produce misleading chromogen, so verify the peroxidase block when DAB colour appears without primary antibody (standard IHC practice). Treat an unexpected compartment or cell population as provisional until morphology and controls support it (standard IHC practice).
Boster reagents
Best PCBP1 / Poly(rC)-binding protein 1 IHC Antibodies
A02636-1 has real IHC data from human paraffin sections (catalog image captions). Its listed reactivity includes human, mouse and rat, while no IF/ICC data are supplied (catalog).
Real IHC data
Anti-PCBP1 Antibody ®
Cat # A02636-1
A02636-1 will render with IHC images from human breast cancer, prostate cancer, rectal cancer and tonsil paraffin sections (catalog image captions). Its application list includes IHC, and its listed reactivity covers human, mouse and rat (catalog).
Which to pick: Choose A02636-1 for human paraffin-section IHC: its captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (catalog image captions). No IF/ICC validation or clone designation is supplied for A02636-1 (catalog). For cross-species work, A02636-1 lists mouse and rat reactivity, but the supplied IHC images show human tissue only (catalog reactivity; catalog 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 several 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 sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02636-1); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms or processed chains are annotated (UniProt); Regulation: Low tissue RNA specificity (HPA tissue RNA). 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.