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CUL4B / Cullin-4B · IHC design guide
Design Immunohistochemistry for CUL4B
Plan CUL4B paraffin-section IHC around its general nuclear tissue pattern (HPA tissue IHC). The catalog antibody lists an IHC dilution of 0.5–1 μg/ml (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CUL4B 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 (HPA tissue IHC); cytoplasmic localization also reported (UniProt) | |
| Staining pattern | General nuclear staining in tissue cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Some cell types, including adipocytes, lack detectable staining (HPA tissue IHC) | |
| Regulation | Expression regulation unreported in record (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope effects are unknown (UniProt) |
Section 1
Recommended CUL4B IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by four published CUL4B IHC protocols (PMC6536945; PMC3351389; PMC7335446; PMC6418142).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA1233); 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-CUL4B, 0.5-1μg/ml (datasheet PA1233) |
| 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 | CUL4B-positive staining in glandular cells of adrenal gland (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 retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen); compare published citrate retrieval where needed (PMC3351389; PMC7335446).
Section 2
What Is the Expected CUL4B Staining Pattern?
CUL4B should show predominantly nuclear staining in tissue sections (HPA: general nuclear expression; Enhanced tissue IHC reliability). UniProt also places CUL4B in the cytoplasm and reports no transmembrane segment (UniProt Q13620: subcellular location and topology). Strong examples include adrenal and epididymal glandular cells; hematopoietic, respiratory epithelial, neuronal, and other listed cells show medium staining (HPA: tissue IHC).
| Distinct nuclear staining in adrenal or epididymal glandular cells, with limited surrounding signal (HPA: High in both cell populations). | This matches the reported tissue pattern and is a useful positive result (HPA: general nuclear expression; Enhanced reliability). Compare cells within the section before scoring intensity: HPA reports different levels across cell types (HPA: tissue IHC). |
| Staining is exclusively cytoplasmic throughout the section, with nuclei consistently clear. | Review this as a compartment mismatch with the general tissue pattern (HPA: general nuclear expression). Cytoplasmic CUL4B is possible (UniProt Q13620: cytoplasm), so the mismatch alone does not prove an artefact; check the positive control and nuclear counterstain (general IHC practice). |
| Strong staining appears in adipocytes or lung alveolar cells, where HPA reports no detection (HPA: tissue IHC). | Cross-reactivity or endogenous detection activity is possible, especially if the signal lacks nuclear definition (general IHC practice; HPA: general nuclear expression). Treat the HPA result as a reference pattern, and check a no-primary control before interpreting an unexpected positive. |
| Color spreads across nuclei, cytoplasm, and tissue spaces without clear cell boundaries. | Diffuse background prevents reliable compartment scoring (general IHC practice). Review blocking, washes, detection reagents, and exposure to chromogen; HPA's nuclear pattern cannot establish CUL4B specificity when the section lacks cellular definition (HPA: general nuclear expression). |
| No nuclear signal appears in adequately preserved adrenal or epididymal glandular cells (HPA: High in both cell populations). | An expected positive control has failed, so a negative test section is inconclusive (general IHC practice). Check the antibody's IHC validation and the staining run; the supplied sources do not identify a CUL4B-specific fixation sensitivity (HPA: antibody validation; UniProt Q13620). |
💡Expected CUL4B appearanceCall a section positive when its relevant cells show discernible nuclear staining, with strong examples in adrenal or epididymal glandular cells (HPA: general nuclear expression; High in these cells); widespread structureless color or strong signal in HPA-undetected cells warrants review (HPA: tissue IHC; general IHC practice).
| Cell population and tissue choice | HPA reports High staining in adrenal and epididymal glandular cells, Medium in several epithelial, hematopoietic, and neuronal populations, and no detection in listed adipocytes and alveolar cells (HPA: tissue IHC). Select and score controls by cell type. |
| Antibody evidence for IHC | HPA011880 has Enhanced IHC validation, while CAB017786 is Supported; the other listed antibodies have no IHC status in this payload (HPA: antibody validation). Match interpretation to the antibody actually used. |
| IF/ICC Q&A: Where should the signal appear? | Mainly in the nucleoplasm (HPA: ICC-IF, enhanced location). This supports a nuclear localisation check, but its cell images are ICC-IF evidence, not an IHC protocol or a tissue-specific intensity standard (HPA: subcellular and tissue IHC). |
| Protein form and topology | CUL4B has no transmembrane segment, signal peptide, or propeptide; its annotated chain spans residues 1–913 (UniProt Q13620: topology and processing). These annotations do not specify an antibody epitope or predict its accessibility in a section. |
| Isoforms and modification | UniProt lists three isoforms and modified residues, including phosphorylation sites (UniProt Q13620: isoforms and modified residues). Without an epitope map for the chosen antibody, their effects on IHC staining cannot be assigned. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in the expected positive control | The staining run or antibody may have failed; adrenal and epididymal glandular cells are reported High (HPA: tissue IHC). | Check the antibody's IHC validation, detection reagents, and run controls before calling test tissue negative (HPA: antibody validation; general IHC practice). |
| Broad haze obscures nuclear boundaries | Background from blocking, washes, or detection can obscure a nuclear pattern (general IHC practice; HPA: general nuclear expression). | Review blocking and wash conditions; compare with a no-primary control and score only cell-defined signal (general IHC practice). |
| Only cytoplasmic staining is visible | This differs from HPA's general nuclear tissue pattern, although UniProt also records cytoplasmic localisation (HPA: tissue IHC; UniProt Q13620). | Confirm the nuclear counterstain and compare an expected positive tissue and the no-primary control before assigning specificity (general IHC practice). |
| An HPA-undetected cell population stains strongly | Unexpected signal could reflect cross-reactivity or endogenous detection activity; HPA reports adipocytes and alveolar cells as Not detected (HPA: tissue IHC; general IHC practice). | Inspect its compartment and the no-primary control, then compare the same antibody with a listed positive cell population (HPA: tissue IHC; general IHC practice). |
| Intensity differs sharply between cell populations | CUL4B staining levels vary by population: HPA lists High, Medium, Low, and Not detected examples (HPA: tissue IHC). | Score the named cell populations separately and compare each with its HPA reference level; avoid treating whole-section darkness as one result (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CUL4B staining
PA1233 · IHCAnti-Cullin 4B antibody, PA1233, IHC(P) IHC(P): Zebrafish Body Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000158290-CUL4B/tissue/cerebral+cortex#imid_3298388
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000158290-CUL4B/tissue/colon#imid_3298306
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000158290-CUL4B/tissue/liver#imid_3298323
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000158290-CUL4B/tissue/kidney#imid_3298316
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA011880; green = CUL4B, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA011880; green = CUL4B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CUL4B IHC & IF
🧪Run adrenal gland first: glandular cells should stain at the High level, with adipose tissue adipocytes as the negative comparison (HPA: adrenal gland, glandular cells High; HPA: adipose tissue, adipocytes Not detected). On the adrenal slide, independently verified CUL4B-negative cells should show background only; the HPA row does not identify a specific adrenal cell type as an internal negative (HPA: adrenal gland, glandular cells High).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CUL4B in U-251MG, U2OS, A-431, KOLF2.1J, HeLa BAC 3316, HeLa , NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary/secondary-only controls, a species- and class-matched isotype control for a monoclonal primary or species-matched nonimmune IgG for a polyclonal primary, and a CUL4B-knockout specimen as a biological negative (standard IHC practice). For adrenal gland chromogenic sections, block endogenous peroxidase and check for intrinsic pigment before scoring staining (standard IHC practice; HPA: adrenal gland, glandular cells High).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected IHC(P) caption does not state a fixative (PA1233 caption). Antigen-retrieval dependence is unreported, so optimize retrieval on serial paraffin sections; the evidence does not establish whether frozen sections or IF would be easier (standard IHC practice; PA1233 caption). In adrenal gland, intrinsic pigment can complicate chromogenic scoring and should be distinguished from specific staining (standard IHC practice; HPA: adrenal gland, glandular cells High).
HPA tissue IHC evidence for CUL4B
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CUL4B IHC Tips
Troubleshoot CUL4B paraffin-section IHC by checking nuclear staining, section processing, controls and cell-specific scoring; the IF entry addresses the separate secondary application.
How should I retrieve CUL4B in paraffin sections when nuclear staining is weak?
Use Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min for CUL4B paraffin-section IHC (page retrieval specification). After cooling, compare a positive-control section processed alongside the test section; adrenal glandular cells or epididymal glandular cells offer reported high staining (HPA: High in both). If nuclei remain weak, test a shorter or longer retrieval exposure on serial sections while holding antibody concentration and detection constant (standard IHC practice). Record nuclear signal, tissue morphology and background together, since stronger retrieval can improve epitope access while also damaging sections or increasing nonspecific staining (standard IHC practice).
Can fixation explain poor CUL4B staining in paraffin sections?
CUL4B-specific fixation sensitivity is unknown from the supplied evidence, so a weak result cannot be assigned to a particular fixative or fixation time. Record the actual fixative and duration, then compare sections processed consistently while keeping Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min constant (page retrieval specification; standard IHC practice). Check preserved nuclear detail and whether an independently processed positive control stains as expected (standard IHC practice). The PA1233 zebrafish body tissue IHC(P) caption does not report a fixative, and its paraffin designation does not establish formalin fixation (PA1233 caption).
Should CUL4B stain nuclei, cytoplasm, or both in tissue IHC?
Expect predominantly nuclear chromogenic staining in many tissue cells, because the tissue profile reports general nuclear expression and the subcellular profile places CUL4B mainly in nucleoplasm (HPA: tissue IHC; HPA: subcellular). CUL4B can also occur in cytoplasm, so assess nuclear and cytoplasmic signal separately rather than treating every extranuclear pixel as artefact (UniProt Q13620: subcellular location). Its reported compartment shift across oocyte stages is context specific and should not become a general scoring rule for tissue sections (UniProt Q13620: subcellular note). If cytoplasmic staining dominates unexpectedly, inspect controls, counterstain alignment and background before assigning biological meaning (standard IHC practice).
Could isoforms or modifications account for inconsistent CUL4B IHC staining?
CUL4B has 3 annotated isoforms and several modified residues, including phosphorylation near positions 49 and 53 (UniProt Q13620: isoforms and modified residues). Check the catalog antibody’s disclosed immunogen or mapped epitope against the isoform sequences before assuming that every section should show the same signal (standard IHC practice). The supplied evidence does not map this antibody’s epitope or show whether fixation or retrieval changes recognition of a particular isoform or modification. Compare serial sections with a second, independently mapped antibody if available, and interpret discordance alongside compartment and cell-type patterns (standard IHC practice; HPA: general nuclear expression).
How should I assess CUL4B by IF alongside paraffin-section IHC?
For the separate IF/ICC workflow, pair CUL4B with a marker for the expected cell population and include single-label controls to check channel bleed-through (standard IF practice). Choose a fluorophore in a channel with low autofluorescence for the specimen, and inspect an unstained control before interpreting faint nuclear signal (standard IF practice). Because CUL4B is nucleoplasmic and has no transmembrane segment, permeabilise fixed cells sufficiently for antibody access to the nuclear epitope; optimise detergent exposure against nuclear morphology (HPA: subcellular; UniProt Q13620: topology; standard IF practice). Do not transfer the PA1233 tissue-IHC caption into an IF fixation claim: it reports IHC(P) zebrafish body tissue and gives no fixative (PA1233 caption).
How can I reduce diffuse or speckled background in CUL4B chromogenic IHC?
First distinguish nuclear signal from diffuse deposit by inspecting a no-primary control and the section edges at the same magnification (standard IHC practice; HPA: general nuclear expression). Block endogenous peroxidase before chromogen development, use a suitable protein block, and wash thoroughly between antibody and detection steps (standard chromogenic IHC practice). If background persists, titrate the primary antibody and shorten chromogen development while retaining the same retrieval conditions in matched sections (standard IHC practice; page retrieval specification). Compare suspected staining with nuclear detail in the counterstain; widespread cytoplasmic haze without coherent nuclei warrants caution because CUL4B is mainly nucleoplasmic (HPA: subcellular).
What is a defensible way to quantify CUL4B staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear staining separately from cytoplasmic staining, consistent with general nuclear tissue expression and mainly nucleoplasmic localisation (HPA: tissue IHC; HPA: subcellular). Report the percentage of positive nuclei or an H-score using preset intensity categories, with the same threshold and imaging settings across sections (standard IHC quantification practice). Normalise positive counts to the number of evaluable cells in that population; for spatial counts, report cells per mm² of viable tissue (standard histology practice). Exclude folds, edges and necrotic areas using documented criteria, and report biological replicates and control performance alongside scores (standard IHC practice).
How do I separate credible CUL4B staining from tissue or detection artefact?
A credible positive pattern has staining in intact nuclei of an expected cell population, with concordant controls; adrenal and epididymal glandular cells are reported high, while adipocytes are not detected (HPA: tissue IHC). Treat strong signal confined to section edges, damaged tissue or necrotic areas as suspect and check a no-primary control for detection background (standard IHC practice). Predominant membrane staining is discordant with CUL4B’s reported nucleoplasmic localisation and lack of a transmembrane segment (HPA: subcellular; UniProt Q13620: topology). If DAB appears without primary antibody, investigate endogenous peroxidase or detection reagents before interpreting the deposit as CUL4B (standard chromogenic IHC practice).
Boster reagents
Best CUL4B / Cullin-4B IHC Antibodies
PA1233 has real paraffin-section IHC data from zebrafish body tissue (PA1233 image caption). Its catalog lists human, mouse, rat, and zebrafish reactivity (PA1233 catalog reactivity).
Real IHC data
Anti-Cullin 4B/CUL4B Antibody ®
Cat # PA1233
PA1233 will render with an IHC(P) image of zebrafish body tissue (PA1233 image caption). Its listed applications are IHC and WB; no IF image is supplied (PA1233 catalog applications; PA1233 IF image alts).
Which to pick: For tissue IHC on paraffin sections, choose PA1233, a rabbit antibody with an IHC(P) image from zebrafish body tissue and a listed IHC dilution of 0.5–1 μg/ml (PA1233 catalog host; PA1233 image caption; PA1233 catalog dilution). There is no supported IF/ICC pick because PA1233 has no listed IF application or IF image (PA1233 catalog applications; PA1233 IF image alts). For cross-species planning, PA1233 lists human, mouse, rat, and zebrafish reactivity; its IHC caption identifies paraffin sections but does not report the fixative (PA1233 catalog reactivity; PA1233 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13620 (CUL4B_HUMAN, Cullin-4B).
- Human Protein Atlas. CUL4B tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CUL4B subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. CUL4B antibody validation summary (4 antibodies).
- CUL4B promotes aggressive phenotypes of HNSCC via the activation of the Wnt/β-catenin signaling pathway. Cancer medicine 2019 — PMC6536945.
- Lack of Cul4b, an E3 ubiquitin ligase component, leads to embryonic lethality and abnormal placental development. PloS one 2012 — PMC3351389.
- Cul4B promotes the progression of ovarian cancer by upregulating the expression of CDK2 and CyclinD1. Journal of ovarian research 2020 — PMC7335446.
- CUL4B promotes prostate cancer progression by forming positive feedback loop with SOX4. Oncogenesis 2019 — PMC6418142.
- PubMed PMID:14578910 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CUL4B (Cullin-4B). Expected localisation Nuclear staining (HPA tissue IHC); cytoplasmic localization also reported (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: PA1233 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA1233); 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-CUL4B, 0.5-1μg/ml (datasheet PA1233); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Some cell types, including adipocytes, lack detectable staining (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope effects are unknown (UniProt); Regulation: Expression regulation unreported in record (UniProt). Evidence: UniProt Q13620, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet PA1233, PMC6536945, PMC3351389. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CUL4B/immunohistochemistry-ihc-guide.