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WEE1 / Wee1-like protein kinase · IHC design guide
Design Immunohistochemistry for WEE1
Plan paraffin-section WEE1 IHC using the observed nuclear staining pattern (HPA tissue IHC). The catalog antibody's tissue caption provides detection conditions (datasheet A01319-1), while WEE1's cell-cycle-dependent abundance informs intensity scoring (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
WEE1 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 several tissues (HPA tissue IHC) | |
| Staining pattern | High nuclear staining in gallbladder glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01319-1) | |
| Positive control | Gallbladder+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 | Cell-cycle stage may alter staining intensity (UniProt) | |
| Regulation | Protein level falls at M/G1 (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage for each (UniProt) |
Section 1
Recommended WEE1 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet A01319-1). Compare its IHC-P protocol with the four published WEE1 IHC protocols below (PMC2663812; PMC3751528; PMC9588842; PMC6090987).
| Sample | Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A01319-1) |
| Fixation | Image fixative and duration unreported (datasheet A01319-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A01319-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01319-1) |
| Primary antibody | Rabbit anti-WEE1, 0.5-1μg/ml (datasheet A01319-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01319-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01319-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | WEE1-positive staining in glandular cells of gallbladder (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01319-1); adjust retrieval to match a published protocol when using that study’s antibody and samples.
Section 2
What Is the Expected WEE1 Staining Pattern?
WEE1 is a nuclear kinase with no transmembrane segment (UniProt P30291: location, topology). In paraffin IHC, expect nuclear staining in selected cells, including gallbladder glandular cells, nasopharyngeal respiratory epithelial cells, placental decidual cells, and testicular preleptotene spermatocytes (HPA: High in each). The HPA tissue profile is Approved, with medium consistency between antibody staining and RNA expression data (HPA: tissue IHC reliability).
| Distinct nuclear chromogen in gallbladder glandular cells or nasopharyngeal respiratory epithelial cells. | This matches the reported compartment and two High staining cell populations (UniProt P30291: nucleus; HPA: High in both). Assess the named cells within the tissue, since a tissue label alone does not identify which cells carry the expected signal (HPA: cell-level tissue IHC profile). |
| Predominantly cytoplasmic or membrane-edge staining, with little nuclear signal. | Treat this as a localization mismatch requiring investigation: WEE1 is annotated as nuclear and lacks a transmembrane segment (UniProt P30291: location, topology). Check the counterstain, detection background, and the IHC-validated antibody’s controls before assigning the chromogen to WEE1 (standard IHC practice). |
| Strong staining in a reported Not detected population, such as adipocytes or prostate glandular cells. | Consider cross-reactivity or endogenous detection activity, especially if expected positive nuclei are unstained (HPA: Not detected in these cells; standard IHC practice). A reported Not detected result is an observed HPA pattern, not proof that every specimen must be negative (HPA: tissue IHC profile). |
| Diffuse chromogen across nuclei, cytoplasm, and tissue spaces without clear cellular boundaries. | This cannot be scored confidently as nuclear WEE1. Nonspecific antibody binding, incomplete blocking, or detection background are possible general IHC causes (standard IHC practice); compare a no-primary control and inspect whether staining follows the expected cell populations (HPA: tissue IHC profile). |
| No nuclear signal in gallbladder glandular cells despite readable morphology and counterstain. | The result conflicts with a reported High population but does not alone establish biological absence (HPA: High in gallbladder glandular cells). Review the catalog antibody’s IHC-P instructions, its recommended dilution, retrieval, and detection controls before interpreting this specimen (standard IHC practice). |
💡Expected WEE1 appearanceCall a result positive when discrete nuclear chromogen is visible in a reported WEE1-positive cell population, with High staining expected in the four named High populations; predominantly cytoplasmic, membrane-edge, or diffuse field-wide color warrants investigation (UniProt P30291: nucleus, topology; HPA: tissue IHC profile; standard IHC practice).
| Compartment and topology | UniProt places WEE1 in the nucleus and reports no transmembrane segment (UniProt P30291: location, topology). Use nuclear localization to judge the IHC pattern; a membrane outline is not the predicted distribution. |
| Cell population and staining level | HPA reports High staining in four named populations, Medium in bone marrow hematopoietic cells and several epithelia, and Not detected in selected other populations (HPA: tissue IHC profile). These are cell-specific observations, not universal scores for whole organs. |
| Cell-cycle context | WEE1 activity rises in S/G2 and falls at M phase; a correlated protein decrease is reported at M/G1 (UniProt P30291: function). Cell-cycle composition may affect interpretation, but the supplied IHC data give no phase-specific staining threshold. |
| Isoforms and modifications | Two isoforms and multiple modified residues are annotated (UniProt P30291: isoforms, modified residues). The supplied sources do not map the IHC antibody’s epitope to either isoform or modification, so staining cannot be assigned to one from this record. |
| IHC evidence and antibody choice | The HPA tissue profile is Approved with medium staining–RNA consistency, and CAB004619 has Approved IHC status (HPA: tissue IHC reliability; HPA: antibody validation). These ratings support a reference pattern, while individual slides still need appropriate controls (standard IHC practice). |
| IF/ICC Q&A: where should signal appear? | HPA reports supported nucleolar localization in ICC-IF; HPA068845 has Supported ICC status, whereas CAB004619 has Approved IHC status (HPA: subcellular; HPA: antibody validation). Do not assume the two assays or antibodies yield identical compartment detail. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue has no nuclear chromogen. | A failed staining or detection step is possible; gallbladder glandular cells are reported High (HPA: tissue IHC; standard IHC practice). | Confirm the cells sampled, then check the catalog antibody’s IHC-P retrieval and dilution instructions, reagent controls, and counterstain readability (standard IHC practice). No WEE1-specific fixation sensitivity is established by the supplied sources. |
| Signal is mainly cytoplasmic or at cell borders. | That distribution conflicts with nuclear localization and the absence of a transmembrane segment (UniProt P30291: location, topology). | Compare a known-positive cell population and a no-primary control; assess whether background or compartment scoring explains the pattern before calling it WEE1 (HPA: tissue IHC; standard IHC practice). |
| Unexpectedly strong staining appears in adipocytes or prostate glandular cells. | These are reported Not detected populations, so cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; standard IHC practice). | Check a no-primary control and inspect nuclear versus diffuse color. If using peroxidase detection, evaluate endogenous peroxidase blocking according to the detection system (standard IHC practice). |
| The entire section has weak, even color. | Diffuse background can obscure cell-specific nuclear staining; nonspecific binding or detection background is possible (standard IHC practice). | Use a no-primary control, review blocking and washes, and compare the expected positive and reported Not detected cell populations on interpretable sections (HPA: tissue IHC; standard IHC practice). |
| Only some nuclei stain in an otherwise positive population. | HPA reports population-level staining, while UniProt describes cell-cycle-dependent WEE1 activity and a protein decrease at M/G1 (HPA: tissue IHC; UniProt P30291: function). | Score the named cell population and nuclear compartment across the section. Do not infer a cell-cycle phase from WEE1 chromogen alone; no phase-specific IHC cutoff is supplied (standard IHC practice). |
| IHC nuclei appear broader than the nucleolar IF/ICC pattern. | The tissue IHC profile describes nuclear expression; separate ICC-IF evidence places WEE1 in nucleoli, using a different validated antibody (HPA: tissue IHC; HPA: subcellular; HPA: antibody validation). | Interpret each assay against its own HPA pattern and antibody validation. For this chromogenic paraffin section, assess nuclear staining in the reported cell populations (HPA: tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing WEE1 staining
A01319-1 · IHCIHC analysis of WEE1 using anti-WEE1 antibody (A01319-1). WEE1 was detected in paraffin-embedded section of human mammary cancer tissue. Hea
A01319-1 · IFIF analysis of WEE1 using anti-WEE1 antibody (A01319-1). WEE1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retri
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000166483-WEE1/tissue/cerebral+cortex#imid_1529600
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000166483-WEE1/tissue/colon#imid_1529518
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000166483-WEE1/tissue/liver#imid_1529535
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000166483-WEE1/tissue/kidney#imid_1529520
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA068845; green = WEE1, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA068845; green = WEE1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for WEE1 IHC & IF
🧪Run gallbladder first and require staining in glandular cells (HPA: High in gallbladder glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the gallbladder slide, treat adjacent cells without specific staining as local background, since the supplied HPA row does not identify a validated internal negative cell type.
Positive control tissue: Gallbladder (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show WEE1 in RT-4, Rh30, U2OS, with annotated localisation: Nucleoli (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a rabbit IgG isotype control matched to the primary antibody, and a WEE1-knockout biological negative where available (selected-SKU caption: rabbit primary antibody). For the caption’s biotin-based DAB detection, quench endogenous peroxidase and assess endogenous biotin background (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state the fixative. The reported paraffin-section staining used heat retrieval in EDTA at pH 8.0, so that retrieval condition is a practical starting point (selected-SKU caption); the evidence does not establish whether frozen sections or IF are easier. Check the gallbladder section for background from endogenous peroxidase or biotin under the reported SABC/DAB detection scheme (selected-SKU caption).
HPA tissue IHC evidence for WEE1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Testis | Preleptotene spermatocytes | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced WEE1 IHC Tips
Troubleshoot WEE1 staining in paraffin sections by checking retrieval, nuclear localisation, cell type and detection controls before comparing scores.
What should I change when WEE1 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A01319-1). The selected tissue image used that retrieval, followed by 1 μg/ml primary antibody overnight at 4°C (caption A01319-1). If staining remains weak, compare a shorter and longer heating cycle on adjacent sections while keeping antibody incubation and detection constant (standard IHC practice). Check whether glandular cells in gallbladder or respiratory epithelial cells in nasopharynx show nuclear signal as positive tissue references (HPA: High in both cell populations; HPA: nuclear expression). Avoid judging retrieval by diffuse cytoplasmic colour alone, because WEE1 is annotated as nuclear (UniProt P30291: subcellular location).
Can fixation explain weak or uneven WEE1 staining in paraffin sections?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A01319-1). Record the fixative, fixation duration, tissue thickness and processing history for each specimen before comparing WEE1 scores (standard IHC practice). When signal varies across a block, compare central and peripheral regions and repeat staining on sections processed together to assess handling effects (standard IHC practice). Keep retrieval at EDTA pH 8.0 during that comparison so fixation is the main variable under review (datasheet A01319-1; standard IHC practice). HPA staining patterns and WEE1 sequence features do not establish how fixation changes this antibody’s epitope (HPA: tissue IHC profile; UniProt P30291: sequence annotation).
Should WEE1 staining be nuclear, nucleolar or cytoplasmic?
Prioritise nuclear staining when reading chromogenic WEE1 sections (UniProt P30291: nucleus; HPA: nuclear expression in several tissues). A nucleolar accent is plausible, although that finer localisation comes from cell imaging rather than this paraffin-section caption (HPA subcellular: supported nucleoli; caption A01319-1). Compare the brown signal with a nuclear counterstain and record nuclear and cytoplasmic staining separately (standard IHC practice). WEE1 has no annotated transmembrane segment, so a crisp membrane-only pattern warrants a specificity check (UniProt P30291: topology). Use a no-primary control and an independently stained reference section when diffuse cytoplasmic colour obscures the nuclear pattern (standard IHC practice).
Could isoforms or phosphorylation change what the antibody detects in tissue?
WEE1 has 2 annotated isoforms and multiple modified residues, including phosphorylation sites near the N terminus (UniProt P30291: isoforms and modified residues). Its protein kinase domain spans residues 299–569, but the selected caption does not identify the antibody epitope (UniProt P30291: domain; caption A01319-1). Therefore, do not interpret a negative section as loss of a particular isoform or phosphorylation state without epitope information and separate validation (standard IHC interpretation). Keep retrieval, primary incubation and chromogen development consistent across specimens when comparing staining (standard IHC practice). If a phenotype depends on isoform or modification status, use an independently validated reagent or assay specific to that feature (standard IHC practice).
How should I adapt WEE1 localisation checks to multiplex immunofluorescence?
Use a marker for the expected cell population alongside WEE1, such as a validated glandular-cell marker in gallbladder, and assess WEE1 within marked cells (HPA: High in gallbladder glandular cells; standard IF practice). Choose fluorophores after checking unstained tissue for autofluorescence, and place the weaker channel where tissue background is lower (standard IF practice). Include a nuclear counterstain to test whether signal is nuclear or nucleolar (UniProt P30291: nucleus; HPA subcellular: supported nucleoli). Because WEE1 lacks a transmembrane segment and the antibody’s epitope is unspecified, optimise permeabilisation for access to intracellular and nuclear epitopes (UniProt P30291: topology; caption A01319-1: epitope not specified; standard IF practice). Validate this IF workflow separately from the paraffin-section IHC conditions (standard IF practice).
How can I distinguish nonspecific brown staining from WEE1 signal?
First inspect a no-primary section for colour from the detection system and tissue itself (standard IHC practice). The selected example uses a biotinylated secondary, streptavidin-biotin complex and DAB, so a detection-only control helps assess background from those steps (caption A01319-1; standard IHC practice). Apply an appropriate peroxidase block before DAB development and assess whether biotin-based detection adds unwanted staining (standard chromogenic IHC practice). Compare suspicious colour with the nuclear pattern expected for WEE1 and with staining in a reference cell population (UniProt P30291: nucleus; HPA: High in gallbladder glandular cells). If diffuse colour persists, adjust blocking, washing or detection development one variable at a time (standard IHC practice).
How should I score WEE1 staining across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear scoring rule before examining outcomes, because WEE1 is annotated as nuclear (UniProt P30291: subcellular location; standard IHC practice). Report the percentage of positive nuclei and an H-score from intensity categories, or positive nuclei per mm² when cell density is the question (standard IHC scoring practice). Normalise counts to the number of evaluable nuclei in the same cell population, and exclude folds, necrosis and poorly preserved areas (standard IHC scoring practice). Keep retrieval at EDTA pH 8.0 and use the same detection settings across compared sections (datasheet A01319-1; standard IHC practice). Record nuclear and cytoplasmic scores separately rather than combining them (UniProt P30291: nucleus; standard IHC scoring practice).
What evidence makes a WEE1-positive cell convincing rather than artefactual?
Look for reproducible nuclear signal in identifiable cells, with a plausible nucleolar accent, rather than colour confined to section edges (UniProt P30291: nucleus; HPA subcellular: supported nucleoli; standard IHC interpretation). High staining in gallbladder glandular cells or nasopharyngeal respiratory epithelial cells provides a reference, while adipocytes are listed as not detected (HPA: tissue IHC profile). Treat staining in necrotic regions, at folds or over tissue edges as suspect until it reproduces in intact areas (standard IHC interpretation). A no-primary control helps identify endogenous enzyme or detection-system colour before calling a cell positive (standard chromogenic IHC practice). Interpret negative or discordant cells cautiously because HPA rates antibody staining versus RNA expression as having medium consistency (HPA: Approved, medium consistency).
Boster reagents
Best WEE1 / Wee1-like protein kinase IHC Antibodies
The catalog antibody has IHC data from human mammary cancer paraffin sections and IF data from U20S cells; listed reactivity covers human, mouse and rat (A01319-1 image captions; catalog reactivity).
Real IHC data
Anti-WEE1 Antibody ®
Cat # A01319-1
A01319-1 will render with its own IHC figure from a human mammary cancer paraffin section (A01319-1 IHC image caption). It is also listed for IF and ICC, with an IF image from U20S cells and stated human, mouse and rat reactivity (A01319-1 IF image caption; catalog applications and reactivity).
Which to pick: For tissue IHC, choose A01319-1: its figure documents a paraffin section with EDTA retrieval at pH 8.0; the fixative is unreported (A01319-1 IHC image caption). For IF/ICC, the same SKU has an IF image from U20S cells and lists both applications (A01319-1 IF image caption; catalog applications). It is also the listed cross-species option for human, mouse and rat, though clonality is unreported (catalog reactivity; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P30291 (WEE1_HUMAN, Wee1-like protein kinase).
- Human Protein Atlas. WEE1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. WEE1 subcellular location (ICC-IF): Localized to the nucleoli..
- Human Protein Atlas. WEE1 antibody validation summary (2 antibodies).
- Integrated functional, gene expression and genomic analysis for the identification of cancer targets. PloS one 2009 — PMC2663812.
- High expression of wee1 is associated with malignancy in vulvar squamous cell carcinoma patients. BMC cancer 2013 — PMC3751528.
- Combined evaluation of both WEE1 and phosphorylated cyclin dependent kinase 1 expressions in oral squamous cell carcinomas predicts cancer recurrence and progression. Journal of dental sciences 2022 — PMC9588842.
- Targeting WEE1 to enhance conventional therapies for acute lymphoblastic leukemia. Journal of hematology & oncology 2018 — PMC6090987.
- PubMed PMID:7743995 — UniProt-cited evidence.
- PubMed PMID:11528126 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for WEE1 (Wee1-like protein kinase). Expected localisation Nuclear staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01319-1). Validated IHC antibody: A01319-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A01319-1); 2. Fixation: Image fixative and duration unreported (datasheet A01319-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A01319-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01319-1); 8. Primary antibody: Rabbit anti-WEE1, 0.5-1μg/ml (datasheet A01319-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01319-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Cell-cycle stage may alter staining intensity (UniProt); Isoform / epitope: 2 isoforms; check epitope coverage for each (UniProt); Regulation: Protein level falls at M/G1 (UniProt). Evidence: UniProt P30291, HPA tissue IHC · Approved, HPA ICC-IF · nucleoli., Datasheet A01319-1, PMC2663812, PMC3751528. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/WEE1/immunohistochemistry-ihc-guide.