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UHRF1 / E3 ubiquitin-protein ligase UHRF1 · IHC design guide
Design Immunohistochemistry for UHRF1
Plan UHRF1 paraffin-section IHC around the nuclear staining observed in thymus (HPA tissue IHC). This guide covers fixation, controls and scoring for the catalog antibody’s IHC-P application (datasheet PB9905).
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 fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | S-phase chromatin binding may shift nuclear pattern (UniProt) | |
| Regulation | Overexpressed in breast cancer (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage needs checking (UniProt) |
Section 1
Recommended UHRF1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: PB9905) is followed by published chromogenic protocols for lung, bladder, and breast 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 EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: PB9905). Use each article’s stated retrieval when reproducing its protocol (PMC2906730; PMC2713709; PMC8797458).
Section 2
What Is the Expected UHRF1 Staining Pattern?
UHRF1 should appear predominantly in nuclei, consistent with its nuclear and chromosome localization and lack of a transmembrane segment (UniProt Q96T88). Expect prominent staining in thymic cortical cells and ovarian follicle cells, both scored High by HPA (HPA tissue IHC). The tissue IHC profile has Enhanced reliability, with high consistency between staining and RNA expression (HPA tissue IHC).
| Distinct nuclear staining in thymic cortical cells, with limited signal outside nuclei. | This matches the reported High staining in thymic cortical cells (HPA tissue IHC) and nuclear localization (UniProt Q96T88). Interpret the cells and compartment together; a dark section alone does not establish a correct UHRF1 pattern. |
| Predominantly cytoplasmic or membranous staining, especially when nuclei remain clear. | Treat this as a possible staining artefact or off-target signal: UHRF1 is assigned to the nucleus and chromosomes and has no transmembrane segment (UniProt Q96T88). Check the counterstain and a known-positive section before scoring it as UHRF1. |
| Strong staining in an HPA-listed cell type scored Not detected, such as adipocytes. | Consider cross-reactivity or endogenous chromogenic activity (general IHC practice). HPA reports UHRF1 as Not detected in adipocytes (HPA tissue IHC), but that observation is a comparison point, not proof that every adipocyte must be negative. |
| Diffuse colour over nuclei, cytoplasm, and surrounding tissue, with poorly defined cell boundaries. | The pattern cannot be assigned confidently to nuclear UHRF1 (UniProt Q96T88). Uneven blocking, residual detection reagents, or excessive detection can produce nonspecific background (general IHC practice); compare with a control lacking primary antibody. |
| No nuclear signal in thymic cortical cells on an otherwise interpretable section. | This conflicts with their High HPA staining and the reported strong nuclear expression in thymus (HPA tissue IHC). Review the positive-control section and assay steps before calling the sample negative; a failed positive control makes absence of signal uninterpretable (general IHC practice). |
💡Expected UHRF1 appearanceCall a result positive when staining is clearly nuclear in the expected cells, especially High thymic cortical or ovarian follicle cells (HPA tissue IHC); broad cytoplasmic or membranous colour is a suspect positive given UHRF1's nuclear localization (UniProt Q96T88).
| Cell type within the section | Expected intensity differs by cell type: thymic cortical and ovarian follicle cells are High, whereas testicular spermatogonia are Medium (HPA tissue IHC). Select and score identifiable cells rather than treating an entire organ as uniformly positive. |
| Nuclear distribution and cell cycle | UHRF1 associates with replicating DNA through S phase, including pericentric heterochromatin; non-S-phase nuclear distribution is described as homogeneous by similarity (UniProt Q96T88). Nuclear texture may vary, so require the correct compartment rather than identical nuclear appearance in every cell. |
| IHC evidence and antibody validation | The tissue profile has Enhanced reliability and reports strong nuclear thymus staining (HPA tissue IHC). Both listed antibodies, HPA049408 and HPA055446, have Enhanced IHC status (HPA antibodies); this supports the reported pattern without guaranteeing every specimen or assay run. |
| Isoforms and epitope coverage | Two UHRF1 isoforms are listed (UniProt Q96T88). The supplied record gives no antibody epitope or isoform coverage, so a staining difference cannot be assigned to a particular isoform from these sources alone. |
| IF/ICC Q: Where should fluorescence appear? | A: Expect nucleoplasmic signal; HPA marks that location Supported and lists images from HEK293, MCF-7, and U2OS (HPA subcellular ICC-IF). This answers localization only; the IF/ICC guide covers its assay workflow. |
| Situation | Likely cause | Next action |
|---|---|---|
| Thymus positive control shows no nuclear staining. | A failed assay step is possible because thymic cortical cells are High and thymus has strong nuclear expression (HPA tissue IHC). | Check section integrity, reagent application, detection, and counterstain with the IHC-validated antibody; withhold negative calls until the positive control works (general IHC practice). |
| Signal is mainly cytoplasmic or membranous. | That distribution disagrees with nuclear and chromosome localization and the absence of a transmembrane segment (UniProt Q96T88). | Verify nuclear boundaries with the counterstain, then compare a known-positive section and a control lacking primary antibody (general IHC practice). |
| Colour appears in cells reported as Not detected by HPA. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); adipocytes are Not detected in the HPA profile (HPA tissue IHC). | Inspect the cell identity and run a control lacking primary antibody before assigning the colour to UHRF1 (general IHC practice). |
| Background obscures nuclear boundaries across the section. | Nonspecific reagent binding or excess detection can obscure localization (general IHC practice). | Review blocking, washes, and detection conditions; compare background with a control lacking primary antibody (general IHC practice). |
| Bone marrow staining looks weak beside thymus. | HPA scores hematopoietic cells Low, while thymic cortical cells are High (HPA tissue IHC). RNA group enrichment in bone marrow does not set the IHC intensity for every cell. | Score the identified cell type against its own HPA reference and confirm that the thymus control shows the expected nuclear pattern (HPA tissue IHC). |
| Nuclei stain, but intensity varies from cell to cell. | UHRF1 associates with replicating DNA during S phase, and its non-S-phase nuclear distribution is described by similarity (UniProt Q96T88); staining variation alone does not identify cell-cycle phase. | Score compartment and cell type first. Use an independent cell-cycle measure if phase-specific interpretation is needed (general IHC practice). |
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 ovary first: follicle cells should show nuclear UHRF1 staining (HPA: High in follicle cells; UniProt Q96T88: nucleus). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the ovary slide, cells without nuclear signal can serve as internal negatives after their identity is checked against the tissue morphology (HPA: High in follicle cells; UniProt Q96T88: nucleus).
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-species- and clonality-matched isotype control, and UHRF1 knockout tissue or a validated immunogen-peptide competition control (standard IHC practice). For the ovary slide, check endogenous peroxidase and biotin background because the reported chromogenic detection uses a biotinylated secondary, streptavidin–biotin complex, and DAB (PB9905 tissue-IHC caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9905 paraffin-section caption does not state the fixative (PB9905 tissue-IHC caption). The caption reports heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required or define an optimal retrieval condition (PB9905 tissue-IHC caption). Whether frozen sections or IF are easier is unreported; in ovary, score follicle-cell nuclei against adjacent tissue morphology and control-slide background (HPA: High in follicle 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 in paraffin sections by checking retrieval, nuclear localisation, cell type and controls before interpreting signal.
How should I optimise antigen retrieval when UHRF1 nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PB9905). The catalog antibody produced staining in a human intestinal cancer paraffin section after that retrieval, followed by 1 μg/ml primary antibody overnight at 4°C (datasheet PB9905). If nuclei remain weak, compare retrieval duration on matched sections while holding antibody concentration and detection constant, and check for tissue damage (standard IHC practice). Include a section with expected nuclear staining, such as thymus cortical cells (HPA: High in thymus cortical cells). Document retrieval duration and nuclear signal together so improved staining is not mistaken for increased background (standard IHC practice).
Could fixation explain absent UHRF1 staining in my paraffin sections?
UHRF1-specific fixation sensitivity is unknown here: the catalog image identifies a paraffin section but does not state its fixative (datasheet PB9905). Record the specimen’s fixative and fixation duration, then compare similarly processed sections before attributing weak staining to fixation (standard IHC practice). Excessive or variable fixation can alter epitope accessibility in tissue sections, so keep retrieval and detection conditions matched during that comparison (standard IHC practice). Use a nuclear-positive control processed alongside the specimen, such as thymus cortical cells, to check whether the staining run worked (HPA: High in thymus cortical cells; standard IHC practice). Do not infer a UHRF1-specific fixation effect from staining patterns alone.
Should diffuse cytoplasmic UHRF1 staining be accepted as positive?
Score nuclear staining first: UHRF1 is reported in the nucleus and on chromosomes, and HPA supports nucleoplasmic localisation (UniProt Q96T88 subcellular; HPA subcellular). In replicating cells, UHRF1 can associate with replicating DNA, including pericentric heterochromatin, so nuclear patterns need not look uniform (UniProt Q96T88 subcellular). Predominantly diffuse cytoplasmic DAB signal deserves a review of morphology, background and detection controls before it is called UHRF1-positive (UniProt Q96T88 subcellular; standard IHC practice). Compare the suspect cells with intact nuclei in the same section and with an expected nuclear-positive control (standard IHC practice; HPA: High in thymus cortical cells). Record nuclear and cytoplasmic staining separately rather than combining them in one score (standard IHC practice).
Can this antibody distinguish UHRF1 isoforms or phosphorylation states in tissue?
The supplied record lists 2 UHRF1 isoforms and several modified residues, including phosphoserines at positions 76, 91 and 95 (UniProt Q96T88 isoforms; UniProt Q96T88 modified residues). No immunogen sequence or isoform-specific validation is supplied for the catalog antibody, so a positive nucleus cannot identify an isoform or phosphorylation state. Compare serial sections with appropriate independent assays if that distinction matters, and report the IHC result as UHRF1 immunoreactivity (standard IHC practice). Keep retrieval and detection consistent across specimens because changes in epitope accessibility can change apparent staining intensity (standard IHC practice). Interpret compartment and cell type before assigning biological meaning to intensity differences (UniProt Q96T88 subcellular; standard IHC practice).
How can IF help investigate an uncertain UHRF1 IHC pattern?
In a separate IF experiment, pair UHRF1 with a validated marker for the cell population being evaluated and a nuclear counterstain, then inspect channels independently before assessing overlap (standard IF practice). Choose spectrally separated fluorophores and assess unstained tissue autofluorescence; a far-red channel can help when shorter wavelengths show substantial background (standard IF practice). UHRF1 is nuclear and has no transmembrane segment, so permeabilise sufficiently for antibody access to nuclear epitopes while preserving nuclear morphology (UniProt Q96T88 subcellular; UniProt Q96T88 topology; standard IF practice). Include single-label controls to check spectral bleed-through and a secondary-only control for nonspecific fluorescence (standard IF practice). HPA supports nucleoplasmic localisation in ICC/IF images from HEK293, MCF-7 and U2OS cells, but those images do not establish performance for this catalog antibody (HPA subcellular).
What causes widespread brown staining despite weak UHRF1 nuclear signal?
First distinguish nuclear DAB signal from staining over vessels, damaged areas and section edges by examining intact cell morphology (standard IHC practice). The catalog example used 10% goat serum blocking, biotinylated goat anti-rabbit secondary antibody and DAB detection, so review each corresponding reagent and control when following that method (datasheet PB9905). Include a no-primary control to assess detection-system background, and block endogenous peroxidase as part of a general chromogenic IHC workflow (standard IHC practice). If the no-primary section stains, optimise the detection steps before increasing primary antibody concentration (standard IHC practice). Judge success by clearer nuclear signal in expected cells, rather than by overall brown intensity (UniProt Q96T88 subcellular; HPA tissue IHC).
How should I quantify heterogeneous UHRF1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis area before scoring, and count intact nuclei within that area rather than all brown pixels (UniProt Q96T88 subcellular; standard IHC practice). Report the percentage of nuclear-positive cells, or use an H-score combining the percentages at intensity grades 0–3 for a 0–300 scale (standard IHC practice). For spatial comparisons, report positive nuclei per mm² alongside total evaluable nuclei per mm², so differences in cell density remain visible (standard IHC practice). Apply the same positivity threshold, retrieval conditions and imaging settings to every section in the comparison (standard IHC practice). Stratify distinct cell populations when needed: HPA reports High staining in thymus cortical cells but no detected staining in adipocytes (HPA tissue IHC).
How can I separate true UHRF1 positivity from staining artefacts?
A convincing result places signal in intact nuclei of an appropriate cell population and is supported by a clean no-primary control (UniProt Q96T88 subcellular; standard IHC practice). Strong nuclear expression in thymus cells is reported, while staining was not detected in adipocytes in the cited HPA tissue profile; use those observations as cell-specific context (HPA tissue IHC). Treat predominant cytoplasmic colour, edge-only signal and staining over necrotic tissue as reasons to re-examine the section and controls (UniProt Q96T88 subcellular; standard IHC practice). Check endogenous peroxidase background when interpreting DAB signal, particularly if colour appears without primary antibody (standard IHC practice). Nuclear staining alone does not demonstrate UHRF1 ligase activity, DNA methylation maintenance or a particular cell-cycle phase (UniProt Q96T88 function; standard IHC practice).
Boster reagents
Best UHRF1 / E3 ubiquitin-protein ligase UHRF1 IHC Antibodies
The catalog includes UHRF1 antibodies with human paraffin-section IHC data (PB9905 image caption) and human cell IF/ICC data (PB9905 and A01156-1 image captions); A01156-1 also lists mouse reactivity (catalog reactivity).
Real IHC data
Anti-UHRF1 Antibody ®
Cat # PB9905
Real IF data
Anti-UHRF1 Antibody ®
Cat # A01156-1
PB9905 has IHC data from a paraffin-embedded human intestinal cancer section and IF/ICC data from U20S cells (PB9905 image captions). A01156-1 has IF/ICC data from MCF-7 cells and lists human and mouse reactivity, with no IHC application listed (A01156-1 image caption; catalog applications and reactivity).
Which to pick: Choose PB9905 for human paraffin-section IHC: its caption reports EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody; the fixative is unreported (PB9905 IHC image caption). For IF/ICC, either SKU has its own cell image, while A01156-1 is the choice when mouse reactivity is needed (PB9905 and A01156-1 IF image captions; A01156-1 catalog reactivity). Both antibodies are rabbit-hosted, and neither catalog entry specifies clonality (catalog host and clone fields).
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 fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: S-phase chromatin binding may shift nuclear pattern (UniProt); Isoform / epitope: 2 isoforms; epitope coverage needs checking (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.