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POLB / DNA polymerase beta · IHC design guide
Design Immunohistochemistry for POLB
Plan chromogenic POLB IHC in paraffin sections around the general nuclear tissue pattern (HPA tissue IHC). Use high-staining glandular cells as positive references and adipocytes as a low-signal comparison (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
POLB 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 | Nuclear staining in tissue; cytoplasmic POLB is also annotated (HPA tissue IHC; UniProt) | |
| Staining pattern | General nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
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 | Adipocytes may lack detectable staining despite the general pattern (HPA tissue IHC) | |
| Regulation | DNA damage drives nuclear translocation (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–335 chain (UniProt) |
Section 1
Recommended POLB IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is followed by four published POLB chromogenic IHC methods (PMC9869049; PMC7957714; PMC13232957; PMC4964131).
| Sample | Paraffin-embedded rat lung tissue; fixative not specified (datasheet A01946-2) |
| Fixation | Image fixative and duration unreported (datasheet A01946-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-POLB, 1:50-1:200 (datasheet A01946-2) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | POLB-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Citrate pH 6 is a published alternative (PMC9869049).
Section 2
What Is the Expected POLB Staining Pattern?
POLB IHC in paraffin sections should show a predominantly nuclear pattern in appropriate cells, including breast glandular cells and bladder urothelial cells (HPA: general nuclear expression; High in both cell types). The tissue IHC profile has Enhanced reliability, reflecting agreement between antibody staining and RNA data (HPA: Enhanced). POLB can also be cytoplasmic under normal conditions and has no transmembrane segment (UniProt P06746: subcellular location; topology).
| Distinct nuclear staining in glandular, urothelial, or squamous epithelial cells. | This fits the reported tissue IHC pattern; breast glandular cells, bladder urothelial cells, and tonsil squamous epithelial cells are High (HPA: tissue IHC). Compare stained cells with neighboring cells rather than treating every nucleus as equally positive (standard IHC practice). |
| Predominantly membrane-edge or extracellular staining, with little nuclear signal. | That distribution does not fit the reported general nuclear IHC pattern or POLB's lack of a transmembrane segment (HPA: tissue IHC; UniProt P06746: topology). Review morphology and controls before calling it specific; cytoplasmic signal alone needs more caution because UniProt also places POLB in the cytoplasm (UniProt P06746: subcellular location). |
| Strong staining in adipocytes, cardiomyocytes, or smooth muscle cells. | Those cell types are reported as Not detected by tissue IHC (HPA: adipose tissue, heart muscle, smooth muscle). Unexpected staining can indicate cross-reactivity or detection activity; compare a no-primary control and the expected nuclear pattern (standard IHC practice). |
| Diffuse color across cells and surrounding tissue, obscuring boundaries. | A uniform haze cannot be scored confidently as cell-specific POLB staining. Check the no-primary control, blocking and detection steps, and whether the counterstain permits nuclear assessment (standard IHC practice); the expected tissue pattern is general nuclear expression (HPA: tissue IHC). |
| No staining in breast glandular cells or bladder urothelial cells. | Both are reported High and can serve as reference-positive cell populations (HPA: breast; urinary bladder). First verify that the relevant cells are present and the detection control worked; then review antibody use and retrieval against its IHC-P instructions (standard IHC practice). |
💡Expected POLB appearanceA convincing positive result has clear, cell-associated nuclear staining in a reported High population such as breast glandular cells, with intensity judged against local background; diffuse haze or isolated membrane outlines are suspect (HPA: general nuclear expression; High in breast glandular cells; UniProt P06746: topology).
| Tissue and cell selection | HPA reports High staining in breast, gallbladder, and salivary gland glandular cells, plus testis round or early spermatids (HPA: tissue IHC). Its Not detected examples include adipocytes and cardiomyocytes (HPA: tissue IHC). Select and interpret controls by cell type, not tissue name alone. |
| Antibody-specific IHC evidence | HPA049104 and CAB011616 have Enhanced IHC status; HPA069013 is Supported (HPA: antibody validation). Validation status helps prioritize an IHC antibody, but a result still needs interpretable morphology and appropriate slide controls (standard IHC practice). |
| Compartment and damage context | UniProt places POLB in both cytoplasm and nucleus and reports nuclear translocation after DNA damage (UniProt P06746: subcellular location). HPA describes general nuclear expression in tissue IHC (HPA: tissue IHC). Do not infer DNA damage from localization in an uncontrolled section. |
| Processing and topology | The annotated POLB chain spans residues 1–335, with no signal peptide, propeptide, or transmembrane segment (UniProt P06746: processing; topology). These annotations support scrutiny of membrane-only staining; they do not establish an antibody epitope or a retrieval condition. |
| Does IF/ICC require the same localization call? | No: HPA reports vesicles as Approved and cytosol as Supported in ICC-IF, while tissue IHC shows general nuclear expression (HPA: subcellular ICC-IF; tissue IHC). Interpret each assay against its own evidence; this IHC section supplies no IF protocol. |
| Retrieval and detection choices | Follow the catalog antibody's IHC-P instructions for retrieval and detection, then assess no-primary and known-positive controls (standard IHC practice). Neither HPA staining levels nor UniProt annotations establish POLB-specific fixation sensitivity or an optimal retrieval setting. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells are blank. | The target cells may be absent from the section, or the IHC workflow may have failed (standard IHC practice); breast glandular cells and bladder urothelial cells are reported High (HPA: tissue IHC). | Confirm cell identity on the counterstained section, check a working positive control, and compare retrieval and antibody use with the IHC-P instructions (standard IHC practice). |
| Signal is mainly cytoplasmic, with weak nuclei. | UniProt reports cytoplasmic POLB under normal conditions, whereas HPA's tissue IHC profile is generally nuclear (UniProt P06746: subcellular location; HPA: tissue IHC). The discrepancy alone cannot identify its cause. | Compare high-staining reference cells, inspect controls, and report the observed compartment explicitly before assigning the result to POLB (standard IHC practice). |
| Unexpected cells stain strongly. | Adipocytes, cardiomyocytes, and smooth muscle cells are Not detected in HPA tissue IHC (HPA: tissue IHC); cross-reactivity or endogenous detection activity may mimic positivity (standard IHC practice). | Check a no-primary control and whether staining is nuclear and cell-associated; treat unexpected staining as unconfirmed until controls support it (standard IHC practice). |
| Brown signal spreads beyond recognizable cells. | Diffuse background can obscure compartment and cell-type assessment (standard IHC practice); it does not match the reported general nuclear pattern (HPA: tissue IHC). | Review blocking, washes, detection exposure, and counterstain with a no-primary control, then score only distinguishable cells (standard IHC practice). |
| A low-staining tissue appears negative. | Neuronal cells in hippocampus and glial cells in caudate are reported Low (HPA: tissue IHC); absence of an obvious chromogenic signal there is less informative than failure in a High reference population. | Assess a reported High population on a comparable run before interpreting a negative result; avoid calling a low-expression sample a failed assay by itself (HPA: tissue IHC; standard IHC practice). |
| IF/ICC images disagree with the IHC slide. | HPA reports vesicle and cytosol localization by ICC-IF but general nuclear expression by tissue IHC (HPA: subcellular ICC-IF; tissue IHC). | Use the assay-specific HPA observations and controls when interpreting each image; do not transfer an ICC-IF localization call directly to paraffin IHC (standard IHC practice). |
Experiment evidence
IHC & IF images showing POLB staining
A01946-2 · IHCImmunohistochemical analysis of paraffin-embedded rat lung tissue using anti-DNA Polymerase beta antibody. Counter stained with hematoxylin.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000070501-POLB/tissue/cerebral+cortex#imid_2972908
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000070501-POLB/tissue/cerebellum#imid_2972895
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000070501-POLB/tissue/colon#imid_2972818
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000070501-POLB/tissue/liver#imid_2972819
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA049104; green = POLB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA049104; green = POLB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for POLB IHC & IF
🧪Run breast first: its glandular cells should stain (HPA: breast glandular cells High). Use adipose tissue as the negative comparator (HPA: adipocytes Not detected); on the breast slide, any unstained cells can serve as internal background references, but the supplied HPA rows identify no specific breast cell type as negative.
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show POLB in A-431, SK-MEL-30, U-251MG, U2OS, THP-1, NIH 3T3, with annotated localisation: Vesicles (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and isotype-matched controls, plus POLB-knockout material as a biological specificity control (standard IHC practice). For chromogenic detection, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A POLB-specific fixation window or fixation effect is unreported, and the selected A01946-2 rat-lung paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). Optimize antigen retrieval empirically; the supplied evidence establishes neither a POLB-specific retrieval requirement nor that frozen sections or IF are easier (selected-SKU caption: paraffin-section IHC; HPA: ICC-IF images). No breast-specific artefact is documented in the supplied evidence.
HPA tissue IHC evidence for POLB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Round or early spermatids | High | Protein (IHC) | HPA → |
| Tonsil | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Section 3
Advanced POLB IHC Tips
Troubleshoot POLB chromogenic IHC in paraffin sections by checking retrieval, compartment, cell type and controls before interpreting staining intensity.
How should I retrieve POLB in paraffin sections when nuclear staining is weak?
For POLB paraffin-section IHC, start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Let sections cool in retrieval buffer, then compare staining with an identically processed positive control and a primary-antibody omission control (standard IHC practice). If staining remains weak, test a shorter or longer heating interval on adjacent sections before considering a different buffer or pH (standard IHC practice). Assess both nuclear signal, which is common in tissue IHC, and cytoplasmic signal, which is also plausible for POLB (HPA: general nuclear expression; UniProt P06746: nucleus and cytoplasm).
Could fixation explain weak or uneven POLB staining?
POLB-specific sensitivity to fixation is unknown from the supplied evidence; the selected image identifies paraffin-embedded rat lung but does not report its fixative (A01946-2 tissue-IHC caption). Record each specimen’s fixative, fixation duration and processing history before comparing staining across blocks (standard IHC practice). On adjacent sections, keep retrieval and antibody incubation constant while checking whether weak areas track tissue thickness, poor morphology or uneven processing (standard IHC practice). Use a well-preserved positive control in the same run, and treat any improvement after retrieval adjustment as a result of that comparison, not proof of a POLB-specific fixation effect (standard IHC practice).
Should POLB staining be nuclear, cytoplasmic or both?
Evaluate nuclear and cytoplasmic staining separately: tissue IHC shows general nuclear expression, while the protein record lists both compartments and notes cytoplasmic localisation under normal conditions (HPA: tissue IHC profile; UniProt P06746: subcellular location). The record also reports movement to the nucleus following DNA damage, so a shift in distribution needs matched biological conditions before interpretation (UniProt P06746: subcellular location). Score identifiable cells rather than assigning diffuse chromogen to a compartment when hematoxylin obscures boundaries (standard IHC practice). If a result appears exclusively in an unexpected compartment, inspect morphology, background controls and retrieval before calling it POLB-specific (standard IHC practice).
How can I assess an apparent epitope-dependent staining difference?
The supplied record annotates one POLB chain spanning residues 1–335 and no isoforms, so an alternate isoform cannot explain a staining difference on this evidence alone (UniProt P06746: processing and isoforms). POLB has no annotated transmembrane segment or glycosylation sites, while acetylation and methylation sites are reported (UniProt P06746: topology and modified residues). Check the catalog antibody’s stated immunogen or epitope, if available, before proposing that a modification masks binding (standard antibody-validation practice). Compare adjacent sections with matched retrieval and controls; agreement between independent epitopes would strengthen assignment, but neither staining difference nor agreement establishes a particular modification (standard IHC practice).
How should an IF follow-up help resolve ambiguous POLB IHC localisation?
Use IF as a separate follow-up to the chromogenic IHC result, with a cell-type marker suited to the sampled tissue and a nuclear counterstain to identify the cells being compared (standard IF practice). For example, glandular cells in breast or gallbladder and urothelial cells in urinary bladder have high tissue-IHC signals (HPA: tissue IHC profile). Choose fluorophores and filter sets that separate the signals from tissue autofluorescence, and include single-label and no-primary controls (standard IF practice). POLB has no transmembrane segment; optimise gentle permeabilisation for intracellular access while checking that cell boundaries and nuclear morphology remain intact (UniProt P06746: topology; standard IF practice).
What should I check when POLB chromogen appears throughout the section?
First compare the section with a primary-antibody omission control and inspect whether chromogen follows tissue edges, folds, pigment or damaged areas (standard IHC practice). Block endogenous peroxidase before DAB detection, then confirm that any residual colour is not present in the omission control (standard chromogenic IHC practice). Titrate the catalog antibody and detection reagent, and adjust blocking or wash conditions using matched sections rather than interpreting a uniformly dark field (standard IHC practice). POLB is reported in nucleus and cytoplasm, but that distribution alone does not establish the identity of diffuse staining without cellular morphology and controls (UniProt P06746: subcellular location; standard IHC practice).
How should I quantify POLB IHC across specimens with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report either an H-score from intensity and percentage of positive cells or the percentage positive within that population (standard IHC scoring practice). Normalise counts to the number of evaluable target cells; if measuring positive-cell density, report cells per mm² of evaluable tissue and state the analysed area (standard IHC practice). Keep section processing, imaging and positivity thresholds consistent across cases, and exclude folds, necrosis and edge artefacts by predefined rules (standard IHC practice). Report nuclear and cytoplasmic scores separately because both locations are recorded for POLB and tissue IHC shows general nuclear expression (UniProt P06746: subcellular location; HPA: tissue IHC profile).
How can I distinguish convincing POLB staining from artefact?
Look for staining within intact, identifiable cells and compare its compartment and cell type with appropriate controls (standard IHC practice). General nuclear tissue staining is reported, with high signal in breast and gallbladder glandular cells and urinary-bladder urothelial cells; POLB can also be cytoplasmic (HPA: tissue IHC profile; UniProt P06746: subcellular location). Treat colour confined to section edges, necrotic regions or poorly preserved tissue as suspect, and check an omission control for endogenous enzyme or detection background (standard IHC practice). A compartment shift may fit the reported response to DNA damage, but chromogenic IHC alone cannot establish that mechanism without matched conditions and independent evidence (UniProt P06746: subcellular location; standard IHC interpretation).
Boster reagents
Best POLB / DNA polymerase beta IHC Antibodies
The catalog includes human, mouse and rat reactive anti-POLB antibodies; A01946-2 also lists zebrafish reactivity (catalog: reactivity). A01946-2 has a rat lung paraffin-section IHC figure (catalog: image caption).
Real IHC data
Anti-DNA Polymerase Beta POLB Antibody
Cat # A01946-2
A01946-2 will render with IHC and ICC listed, human, mouse, rat and zebrafish reactivity, and a rat lung paraffin-section IHC figure (catalog: applications, reactivity and image caption). M01946 will render with IHC and IF/ICC listed and human, mouse and rat reactivity; no IHC or IF figure is supplied (catalog: applications, reactivity and image captions).
Which to pick: Choose A01946-2 for tissue IHC when a matching example matters: its own figure shows a hematoxylin-counterstained rat lung paraffin section, but the fixative is unreported (catalog: A01946-2 image caption). Choose monoclonal M01946 for IF/ICC because IF is explicitly listed, although no IF figure is supplied (catalog: M01946 title, applications and image captions). Choose A01946-2 when zebrafish reactivity is needed; both SKUs list human, mouse and rat reactivity (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P06746 (DPOLB_HUMAN, DNA polymerase beta).
- Human Protein Atlas. POLB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. POLB subcellular location (ICC-IF): Localized to vesicles and cytosol..
- Human Protein Atlas. POLB antibody validation summary (4 antibodies).
- Huwe1 supports B-cell development, B-cell-dependent immunity, somatic hypermutation and class switch recombination by regulating proliferation. Frontiers in immunology 2022 — PMC9869049.
- Silencing DNA Polymerase β Induces Aneuploidy as a Biomarker of Poor Prognosis in Oral Squamous Cell Cancer. International journal of molecular sciences 2021 — PMC7957714.
- Integration of single-cell and bulk RNA sequencing reveals programmed cell death-associated transcriptional programs in sepsis-induced acute lung injury. PloS one 2026 — PMC13232957.
- A Point Mutation in DNA Polymerase β (POLB) Gene Is Associated with Increased Progesterone Receptor (PR) Expression and Intraperitoneal Metastasis in Gastric Cancer. Journal of Cancer 2016 — PMC4964131.
- PubMed PMID:10648175 — UniProt-cited evidence.
- PubMed PMID:7705833 — UniProt-cited evidence.
- PubMed PMID:7914364 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for POLB (DNA polymerase beta). Expected localisation Nuclear staining in tissue; cytoplasmic POLB is also annotated (HPA tissue IHC; UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A01946-2 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat lung tissue; fixative not specified (datasheet A01946-2); 2. Fixation: Image fixative and duration unreported (datasheet A01946-2); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-POLB, 1:50-1:200 (datasheet A01946-2); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Adipocytes may lack detectable staining despite the general pattern (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–335 chain (UniProt); Regulation: DNA damage drives nuclear translocation (UniProt). Evidence: UniProt P06746, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles and cytosol., Datasheet A01946-2, PMC9869049, PMC7957714. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/POLB/immunohistochemistry-ihc-guide.