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UHRF1 / E3 ubiquitin-protein ligase UHRF1 · IHC design guide
Design Immunohistochemistry for UHRF1
Plan paraffin-section UHRF1 IHC around nuclear staining in thymus (HPA tissue IHC). Start with the IHC-validated antibody PB9905 at 0.5–1 μg/mL (datasheet PB9905), then score nuclear signal by cell type and intensity (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UHRF1 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 thymus (HPA tissue IHC) | |
| Staining pattern | Strong nuclear staining in thymic cortical cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9905) | |
| Positive control | Ovary+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 tissue fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Cell-cycle phase may shift the nuclear pattern (UniProt) | |
| Regulation | Overexpressed in breast cancer (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended UHRF1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: PB9905) is accompanied by three published UHRF1 IHC protocols for paraffin sections (PMC2906730; PMC2713709; PMC8797458).
| Sample | Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet PB9905) |
| Fixation | Image fixative and duration unreported (datasheet PB9905); 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 PB9905); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9905) |
| Primary antibody | Rabbit anti-UHRF1, 0.5-1μg/ml (datasheet PB9905) |
| Primary incubation | Overnight at 4 °C (datasheet PB9905) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9905) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UHRF1-positive staining in follicle cells of ovary (HPA tissue IHC: High). HPA tissue profile: Strong nuclear expression in cells in thymus. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: PB9905); published alternatives use pH 9 or citrate pH 6 retrieval (PMC2906730; PMC8797458).
Section 2
What Is the Expected UHRF1 Staining Pattern?
UHRF1 should appear predominantly in nuclei on paraffin-section IHC: thymic cortical cells and ovarian follicle cells show high staining, while several other listed cell populations show medium or low staining (HPA tissue IHC). HPA rates the tissue pattern Enhanced for consistency with RNA expression (HPA tissue IHC). Nuclear and chromosome localization, with no transmembrane segment, supports a nuclear interpretation (UniProt Q96T88 localization and topology).
| Distinct nuclear staining in thymic cortical cells or ovarian follicle cells. | This fits the high-staining populations in the tissue record (HPA tissue IHC: High). Assess the stained cells and compartment together; strong color elsewhere in the section does not establish the same expected pattern (HPA tissue IHC; UniProt Q96T88 localization). |
| Nuclear staining is present but uneven across cells or regions. | Variation can be compatible with the record: lymph-node non-germinal-center cells, placental cytotrophoblasts, testicular spermatogonia and tonsillar germinal-center cells are listed at medium levels (HPA tissue IHC). UHRF1 associates with replicating DNA during S phase and is more homogeneous in non-S-phase nuclei (UniProt Q96T88 localization). |
| Predominantly cytoplasmic, membranous or extracellular color replaces nuclear staining. | Treat this as discordant with the expected compartment (UniProt Q96T88 localization; HPA subcellular: nucleoplasm supported). Review morphology, counterstain and controls before interpreting it as UHRF1; the record reports no transmembrane segment or signal peptide (UniProt Q96T88 topology and processing). |
| Strong staining appears in a cell type listed as not detected, such as adipocytes or bronchial respiratory epithelial cells. | This conflicts with those specific reference observations (HPA tissue IHC: Not detected). Cross-reactivity or endogenous detection activity is possible under general IHC practice; compare a no-primary control and the expected nuclear signal before assigning specificity. The HPA observation alone cannot identify the cause. |
| No discernible nuclear signal appears in thymic cortical cells. | This conflicts with the high thymic cortical-cell reference pattern (HPA tissue IHC: High). Check tissue identity, morphology and assay controls, then review the antibody's IHC-P instructions and detection steps (general IHC practice). A negative slide alone cannot distinguish absent antigen from an assay failure. |
💡Expected UHRF1 appearanceCall a result positive when staining is chiefly nuclear in the relevant cells, especially high in thymic cortical or ovarian follicle cells; isolated cytoplasmic color or strong staining in HPA-listed not-detected cells warrants control review (HPA tissue IHC; UniProt Q96T88 localization).
| Cell and tissue context | The reference is cell-specific: cortical cells in thymus and follicle cells in ovary are high, while bone-marrow hematopoietic cells are low (HPA tissue IHC). Do not score an entire tissue by a single expected intensity. |
| Nuclear distribution and cell cycle | UHRF1 associates with replicating DNA from early to late S phase, including pericentric heterochromatin; non-S-phase nuclear distribution is described as homogeneous by similarity (UniProt Q96T88 localization). Nuclear texture alone is therefore a weak exclusion criterion. |
| Antibody validation | Two listed rabbit polyclonal antibodies, HPA049408 and HPA055446, carry Enhanced IHC status; only HPA049408 has Supported ICC status (HPA antibodies). These labels support the reported applications, but do not validate an unspecified catalog antibody. |
| Target organization | UniProt lists a single 1–793 chain, no signal peptide, no propeptide, no transmembrane segment and two isoforms (UniProt Q96T88 processing, topology and isoforms). Epitope coverage across isoforms cannot be inferred without antibody-specific epitope information. |
| Situation | Likely cause | Next action |
|---|---|---|
| Thymus shows no nuclear signal in cortical cells. | The result differs from the high reference pattern (HPA tissue IHC); tissue sampling or assay performance may need review (general IHC practice). | Confirm cortical-cell morphology, inspect positive and no-primary controls, and follow the IHC-validated antibody's documented IHC-P conditions (general IHC practice). Do not assign a UHRF1-specific fixation effect; none is supplied. |
| The section has widespread haze that obscures nuclei. | Background from blocking, washing or chromogenic detection can obscure localization (general IHC practice); the expected compartment is nuclear (UniProt Q96T88 localization). | Compare the no-primary control, review blocking and washes, and assess whether the counterstain still resolves nuclei (general IHC practice). Score only separable nuclear staining in the relevant cells (HPA tissue IHC). |
| Color is mainly cytoplasmic or outlines cell membranes. | That compartment conflicts with supported nucleoplasmic localization and nuclear tissue staining (HPA subcellular; HPA tissue IHC). The slide cannot identify the cause by appearance alone. | Check the no-primary control and morphology, then review antibody application validation and detection settings (general IHC practice; HPA antibodies for the listed antibodies). |
| Strong color occurs in adipocytes or bronchial respiratory epithelial cells. | Both are listed as not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice), but neither is proven by this observation. | Compare matched no-primary and positive-tissue controls; judge whether signal is nuclear and cell-specific (general IHC practice; UniProt Q96T88 localization; HPA tissue IHC). |
| Bone marrow looks weak compared with thymus. | Low staining in bone-marrow hematopoietic cells and high staining in thymic cortical cells are both recorded (HPA tissue IHC). Their different intensities alone do not indicate an assay fault. | Use thymic cortical cells as the stronger reference and score marrow hematopoietic cells against their own low reference level (HPA tissue IHC). Confirm nuclear localization before interpreting either signal (UniProt Q96T88 localization). |
| Q: Should an IF/ICC image show the same compartment? | Supported nucleoplasmic localization is reported for ICC-IF, with images in HEK293, MCF-7 and U2OS (HPA subcellular). This establishes a localization reference, not an IF/ICC protocol. | Expect nucleoplasmic signal when interpreting IF/ICC images (HPA subcellular). For this IHC-P guide, use the tissue IHC pattern for slide interpretation; IF/ICC methods belong to the separate guide. |
Experiment evidence
IHC & IF images showing UHRF1 staining
PB9905 · IHCIHC analysis of UHRF1 using anti-UHRF1 antibody (PB9905). UHRF1 was detected in a paraffin-embedded section of human intestinal cancer tissu
PB9905 · IFIF analysis of UHRF1 using anti-UHRF1 antibody (PB9905). UHRF1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retr
A01156-1 · IFIF analysis of UHRF1 using anti-UHRF1 antibody (A01156-1) and anti-Tubulin Alpha antibody (M03989-3). UHRF1 was detected in immunocytochemic
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000276043-UHRF1/tissue/cerebral+cortex#imid_20480603
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000276043-UHRF1/tissue/colon#imid_20480089
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000276043-UHRF1/tissue/liver#imid_20480098
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000276043-UHRF1/tissue/kidney#imid_20480083
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA049408; green = UHRF1, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA049408; green = UHRF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UHRF1 IHC & IF
🧪Run thymus first: cortical cells should show nuclear staining (HPA: High in thymus cortical cells; UniProt Q96T88: nucleus). Use adipose tissue as the negative tissue, assessing adipocytes (HPA: Not detected in adipocytes); cells without specific staining within the thymus section should show only counterstain, but the supplied HPA rows do not establish a thymic cell type as an internal negative (HPA: thymus cortical cells High).
Positive control tissue: Ovary (Follicle cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show UHRF1 in HEK293, MCF-7, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a host- and immunoglobulin-class-matched rabbit IgG control for the catalog antibody (datasheet: rabbit anti-UHRF1); and a UHRF1 knockout biological negative where available. Block endogenous peroxidase and check endogenous biotin when using the caption’s biotinylated secondary and SABC–DAB detection (datasheet: PB9905 tissue-IHC caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact PB9905 tissue-IHC caption does not state a fixative (datasheet: PB9905 tissue-IHC caption). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; the supplied evidence also does not establish whether frozen sections or IF are easier (datasheet: PB9905 tissue-IHC caption; HPA: ICC-IF images available). In densely cellular thymus, use morphology and a counterstain to assign nuclear chromogen to cortical cells (HPA: High in thymus cortical cells; UniProt Q96T88: nucleus).
HPA tissue IHC evidence for UHRF1
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 |
|---|---|---|---|---|
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Thymus | Cortical cells | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Placenta | Cytotrophoblasts | Medium | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UHRF1 IHC Tips
Troubleshoot UHRF1 staining by checking nuclear localisation, tissue controls and the processing conditions documented for the catalog antibody.
How should I adjust retrieval when UHRF1 nuclear staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet PB9905). The catalog antibody produced a tissue-IHC image using this retrieval before overnight incubation at 4°C with 1 μg/ml primary antibody (datasheet PB9905). If staining is weak, first confirm that the retrieval solution reaches the intended pH and that sections remain covered during heating and cooling (standard IHC practice). Compare a known positive section, such as thymus cortical cells, in the same run (HPA: High in thymus cortical cells). Increase retrieval exposure cautiously only after checking section adhesion and morphology, because tissue damage can complicate nuclear scoring (standard IHC practice).
Could fixation explain absent UHRF1 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected tissue-IHC caption describes a paraffin section but does not state its fixative (datasheet PB9905). Record the fixative, fixation duration and specimen thickness for each sample, then compare samples processed in the same batch (standard IHC practice). If nuclei are weak across the slide, run a documented positive control and verify EDTA pH 8.0 retrieval before changing primary-antibody concentration (HPA: High in thymus cortical cells; datasheet PB9905). If only one specimen fails, inspect tissue preservation and staining controls before attributing the result to UHRF1 biology (standard IHC practice). Do not infer fixation tolerance from expression patterns or protein features (datasheet PB9905: fixative unreported).
Should UHRF1 staining be diffuse or punctate within nuclei?
Score UHRF1 primarily within nuclei: it is assigned to the nucleus and chromosomes, with supported nucleoplasmic localisation (UniProt Q96T88 subcellular location; HPA: supported nucleoplasm). In replicating cells, signal can associate with DNA from early through late S phase, including pericentric heterochromatin; UHRF1 also occurs in euchromatic regions (UniProt Q96T88 subcellular location). More homogeneous nuclear distribution is described outside S phase by similarity, so a diffuse pattern alone does not invalidate staining (UniProt Q96T88 subcellular location). Compare nuclear DAB with a light counterstain and inspect several fields before scoring apparent puncta (standard IHC practice). Predominantly cytoplasmic staining warrants review of background and antibody specificity (UniProt Q96T88 subcellular location; standard IHC practice).
Could isoforms or epitope masking change the UHRF1 staining pattern?
UHRF1 has 2 listed isoforms, but the supplied caption does not identify the PB9905 epitope or establish isoform-specific detection (UniProt Q96T88 isoforms; datasheet PB9905). Check the antibody’s documented immunogen or epitope before interpreting differences between specimens as isoform changes (standard IHC practice). UHRF1 includes an N-terminal ubiquitin-like region at residues 1–78 and a YDG domain at 419–582, but those coordinates do not locate this antibody’s binding site (UniProt Q96T88 domains; datasheet PB9905). Its listed phosphorylated residues likewise do not establish whether staining depends on phosphorylation (UniProt Q96T88 modified residues). Compare retrieval conditions and a positive control before assigning weak staining to an isoform or modification (standard IHC practice).
How can IF help investigate uncertain UHRF1 IHC localisation?
Use IF/ICC as a separate assay to examine nuclear localisation; the supplied PB9905 tissue image documents chromogenic paraffin-section IHC, not an IF protocol (datasheet PB9905). Multiplex UHRF1 with a marker for the expected cell population, choosing the marker from the specimen’s biology and confirming each antibody separately (standard IF practice). Select a fluorophore channel with low tissue autofluorescence and include an unstained section to assess background in that channel (standard IF practice). Because UHRF1 is nuclear and has no transmembrane segment, permeabilisation must allow antibody access to nuclear epitopes (UniProt Q96T88 subcellular location and topology; standard IF practice). Compare signal with a nuclear counterstain and use single-stain controls when assessing overlap (standard IF practice).
What should I check when DAB obscures nuclear UHRF1 staining?
First compare a section processed without primary antibody to assess background from the detection system (standard IHC practice). The selected image used a biotinylated secondary antibody, a streptavidin–biotin complex and DAB, so background may arise at several detection steps (datasheet PB9905; standard IHC practice). Include a peroxidase block, assess endogenous biotin when using a biotin-based system, and check whether DAB development is excessive (standard IHC practice). The documented tissue section was blocked with 10% goat serum before primary-antibody incubation (datasheet PB9905). If background persists, compare matched sections with adjusted blocking or development conditions while retaining the same positive control and nuclear scoring rule (standard IHC practice).
How should I quantify UHRF1 in heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of nuclei with UHRF1 staining and nuclear intensity (standard IHC practice; UniProt Q96T88 subcellular location). An H-score combines the percentage at each intensity level; report its scoring scale and reader criteria with the result (standard IHC practice). For spatial questions, report positive nuclei per mm² alongside total nuclei per mm², using the same region-selection rules across specimens (standard IHC practice). Normalise positive counts to eligible, intact nuclei rather than total tissue area alone (standard IHC practice). Analyse comparable cell populations separately because HPA reports levels ranging from High in thymus cortical cells to Not detected in adipocytes (HPA: tissue IHC).
How do I distinguish genuine UHRF1 positivity from staining artefacts?
A credible positive signal is predominantly nuclear and occurs in an appropriate cell population, such as thymus cortical cells or ovarian follicle cells (UniProt Q96T88 subcellular location; HPA: High in thymus cortical cells and ovarian follicle cells). Compare suspicious staining with a matched section lacking primary antibody and with a positive-control section (standard IHC practice). Diffuse cytoplasmic colour, staining concentrated at section edges or necrotic regions, and colour persisting without primary antibody warrant investigation before scoring (UniProt Q96T88 subcellular location; standard IHC practice). Check peroxidase blocking and detection background when DAB appears in unexpected compartments (standard IHC practice). A negative result in one cell population should be interpreted against its own expected expression and tissue preservation (HPA: tissue IHC; standard IHC practice).
Boster reagents
Best UHRF1 / E3 ubiquitin-protein ligase UHRF1 IHC Antibodies
Anti-UHRF1 antibodies have illustrated IHC in human paraffin-embedded intestinal cancer tissue (PB9905 IHC caption) and IF/ICC in MCF-7 cells (A01156-1 IF caption); A01156-1 also lists mouse reactivity (catalog).
Real IHC data
Anti-UHRF1 Antibody ®
Cat # PB9905
Real IF data
Anti-UHRF1 Antibody ®
Cat # A01156-1
PB9905 has an IHC image from a human paraffin-embedded intestinal cancer section and lists IHC and IF/ICC applications (PB9905 IHC caption; catalog). A01156-1 has an IF image from MCF-7 cells, lists IF/ICC applications, and lists human and mouse reactivity (A01156-1 IF caption; catalog).
Which to pick: Choose PB9905 for human paraffin-section IHC: its caption documents EDTA retrieval at pH 8.0 and primary antibody at 1 μg/ml (PB9905 IHC caption). For IF/ICC, A01156-1 has an MCF-7 image at 5 μg/ml and is the option with listed mouse reactivity, although that image documents human cells (A01156-1 IF caption; catalog). Neither entry reports a clone (catalog), and the PB9905 paraffin-section caption does not report the fixative (PB9905 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96T88 (UHRF1_HUMAN, E3 ubiquitin-protein ligase UHRF1).
- Human Protein Atlas. UHRF1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. UHRF1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. UHRF1 antibody validation summary (2 antibodies).
- UHRF1 is a novel diagnostic marker of lung cancer. British journal of cancer 2010 — PMC2906730.
- UHRF1 is a novel molecular marker for diagnosis and the prognosis of bladder cancer. British journal of cancer 2009 — PMC2713709.
- Overexpression of UHRF1 and its potential role in the development of invasive ductal breast cancer validated by integrative bioinformatics and immunohistochemistry analyses. Translational cancer research 2019 — PMC8797458.
- Regulation of UHRF1 by microRNA-9 modulates colorectal cancer cell proliferation and apoptosis. Cancer science 2015 — PMC4520634.
- PubMed PMID:10646863 — UniProt-cited evidence.
- PubMed PMID:17067204 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UHRF1 (E3 ubiquitin-protein ligase UHRF1). Expected localisation Nuclear staining in thymus (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9905). Validated IHC antibody: PB9905 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Ovary (Follicle cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet PB9905); 2. Fixation: Image fixative and duration unreported (datasheet PB9905); 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 PB9905); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9905); 8. Primary antibody: Rabbit anti-UHRF1, 0.5-1μg/ml (datasheet PB9905); 9. Primary incubation: Overnight at 4 °C (datasheet PB9905). Watch for: Fixation: Keep tissue fixation consistent (standard IHC practice; not target-specific); Caveat: Cell-cycle phase may shift the nuclear pattern (UniProt); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: Overexpressed in breast cancer (UniProt). Evidence: UniProt Q96T88, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet PB9905, PMC2906730, PMC2713709. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UHRF1/immunohistochemistry-ihc-guide.