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WDHD1 / WD repeat and HMG-box DNA-binding protein 1 · IHC design guide
Design Immunohistochemistry for WDHD1
Use this guide to plan paraffin-section WDHD1 IHC around its broad nuclear staining (HPA tissue IHC). Lymph node germinal center cells show high staining, and the catalog antibody has an IHC dilution range of 1:100–1:300 (HPA tissue IHC; datasheet A06240).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
WDHD1 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 | Broad nuclear staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across many tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A06240) | |
| Positive control | Appendix+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 | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended WDHD1 IHC & IF Protocols
The catalog antibody protocol uses Tris-EDTA pH 9.0 heat retrieval (datasheet A06240). The published IHC protocols below describe nasopharyngeal and breast tumour tissue microarrays (PMC9811654; PMC7680459).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A06240) |
| Fixation | Image fixative and duration unreported (datasheet A06240); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A06240); 20 min, 95–100 °C (standard) |
| 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-WDHD1, 1:100-1:300 (datasheet A06240) |
| 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 | WDHD1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 heat retrieval (datasheet A06240); the nasopharyngeal study specifies EDTA without a pH (PMC9811654), and the breast study omits retrieval conditions (PMC7680459).
Section 2
What Is the Expected WDHD1 Staining Pattern?
WDHD1 is a nuclear protein localized to the nucleoplasm, with no transmembrane segment (UniProt O75717). In paraffin section IHC, expect nuclear staining across multiple tissues, including high staining in appendix and rectal glandular cells, endometrial stromal cells, and lymph node germinal center cells (HPA tissue IHC). HPA describes the overall pattern as ubiquitous nuclear expression, while reporting some cell types as undetected; its tissue IHC reliability is Enhanced, with medium consistency between staining and RNA data (HPA tissue IHC).
| Distinct nuclear staining in appendix or rectal glandular cells, endometrial stromal cells, or lymph node germinal center cells. | This matches cell types scored High in HPA tissue IHC and the nucleoplasmic location assigned by UniProt O75717. Compare nuclei with neighboring cells on the same section; an HPA High designation does not mean every cell in that tissue must stain equally. |
| Signal is predominantly cytoplasmic, membranous, or extracellular, with little nuclear signal. | That distribution conflicts with WDHD1 nucleoplasmic localization and its lack of a transmembrane segment (UniProt O75717; HPA subcellular ICC-IF). Treat it as a questionable IHC result and reassess staining specificity and detection before assigning it to WDHD1. |
| Strong staining appears in adipocytes, cardiomyocytes, or glial cells in caudate or hippocampus. | HPA scores those specific cell populations Not detected in the listed tissues (HPA tissue IHC). Check cell identity and nuclear localization; persistent signal could reflect antibody cross-reactivity or endogenous detection activity (general IHC practice). These scores do not make the entire tissues absolute negative controls. |
| Broad color covers nuclei and surrounding tissue, obscuring cell boundaries. | A diffuse deposit cannot establish the nucleoplasmic pattern reported for WDHD1 (UniProt O75717; HPA subcellular ICC-IF). Excess antibody, incomplete blocking, or detection background are possible general IHC causes; inspect a no-primary control before scoring cells. |
| No nuclear signal appears in glandular cells of an appendix or rectum section selected as a positive comparator. | Both cell populations are scored High by HPA tissue IHC, so a blank result warrants a workflow check. Confirm tissue preservation and that a general nuclear counterstain shows identifiable cells, then review the validated IHC antibody and detection steps (general IHC practice). |
💡Expected WDHD1 appearanceCall positive a predominantly nuclear signal in the specified HPA High cell populations, with cell to cell intensity variation; predominantly cytoplasmic or extracellular color is inconsistent with the reported nucleoplasmic location (HPA tissue IHC; UniProt O75717).
| Compartment and tissue context | UniProt places WDHD1 in the nucleoplasm, and HPA reports a ubiquitous nuclear profile (UniProt O75717; HPA tissue IHC). HPA also scores particular adipocytes, cardiomyocytes, and glial populations Not detected, so interpret staining by identified cell type rather than assuming uniform positivity throughout a section. |
| IHC antibody validation | HPA lists HPA001122 as IHC Enhanced; HPA000632 has ICC Enhanced but no listed IHC validation (HPA antibodies). The tissue profile has Enhanced reliability with medium staining–RNA consistency (HPA tissue IHC). These are validation summaries, not guarantees that every section or detection setup will reproduce the pattern. |
| Isoforms, modifications, and epitope limits | UniProt lists 2 isoforms, 15 modified residues, and one annotated chain spanning residues 1–1129 (UniProt O75717). Without an epitope location in the supplied record, their effects on this antibody’s IHC staining cannot be predicted; do not infer isoform-specific staining or a processing-dependent pattern. |
| Antigen retrieval | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| IF/ICC cross-check — where should fluorescence appear? | In the nucleoplasm: HPA reports an enhanced nucleoplasmic ICC-IF location and lists images from A-431, U-251MG, and U2OS cells (HPA subcellular ICC-IF). That supports a localization cross-check; it does not supply an IF protocol here or transfer ICC validation to the IHC antibody. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known HPA High cell population is blank. | A missed detection step, unsuitable antibody condition, or absent identifiable cells may explain a blank section (general IHC practice). HPA High is an observed tissue score, not a guaranteed result in every preparation (HPA tissue IHC). | Check the IHC-validated antibody identity, section integrity, counterstain, and detection controls; compare with an appendix or rectum section containing the specified glandular cells (HPA antibodies; HPA tissue IHC). |
| The strongest signal lies outside nuclei. | The distribution conflicts with the reported nucleoplasmic location (UniProt O75717; HPA subcellular ICC-IF). Background deposition or cross-reactivity is possible (general IHC practice). | Check nuclear counterstain alignment and a no-primary control, then reassess whether any specific nuclear staining remains (general IHC practice). |
| High color appears in a cell population HPA scores Not detected. | Cell identification may be uncertain, or signal may reflect cross-reactivity or endogenous detection activity (general IHC practice). HPA reports specific negative cell populations, including adipocytes and cardiomyocytes (HPA tissue IHC). | Identify the stained cells morphologically, verify their compartment, and compare no-primary and detection controls before calling them WDHD1 positive (general IHC practice). |
| The entire section has diffuse chromogen. | Diffuse deposition can mask the expected nuclear pattern (UniProt O75717). Excess antibody or insufficient blocking or washing are general IHC possibilities, not WDHD1-specific findings (general IHC practice). | Use a no-primary control; review blocking, washing, antibody concentration, and detection exposure while retaining a known HPA High cell population for comparison (general IHC practice; HPA tissue IHC). |
| Signal differs between lymphoid tissue and other tissues. | HPA reports tissue-enhanced RNA in lymphoid tissue, but its protein IHC profile is described as ubiquitous nuclear with variable cell-level scores (HPA tissue IHC). RNA enrichment alone cannot predict a section’s chromogen intensity. | Score nuclear staining in the named cell populations and record cell identity and intensity separately; compare with the HPA tissue IHC scores rather than ranking whole sections by RNA specificity. |
| ICC-IF appears nucleoplasmic but paraffin IHC is equivocal. | HPA’s enhanced ICC-IF localization and IHC antibody validation apply to different listed antibodies: HPA000632 and HPA001122, respectively (HPA antibodies; HPA subcellular ICC-IF). | Use the ICC-IF finding as a compartment check, then judge paraffin IHC against the IHC antibody’s validation and HPA cell-specific tissue observations (HPA antibodies; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing WDHD1 staining
A06240 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA, pH9.0 was used for antigen retrieval. 2 Antibody was diluted a
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198554-WDHD1/tissue/cerebral+cortex#imid_528338
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000198554-WDHD1/tissue/stomach+1#imid_528238
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198554-WDHD1/tissue/colon#imid_528256
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198554-WDHD1/tissue/liver#imid_528265
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA000632; green = WDHD1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA000632; green = WDHD1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for WDHD1 IHC & IF
🧪Run lymph node first: germinal center cells should show nuclear staining (HPA: High in lymph-node germinal center cells; UniProt: nucleoplasm). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the positive slide, cells without nuclear signal provide a background reference, although the supplied HPA rows do not establish a specific internal negative cell type.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show WDHD1 in A-431, U-251MG, U2OS, U2OS, siRNA 2 (10x), U2OS, siRNA 2 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched isotype control matching the primary antibody’s host species and clonality, and a WDHD1-knockout sample or immunizing-peptide block where available (standard IHC control practice). Quench endogenous peroxidase and inspect lymph-node sections for residual background before scoring chromogenic staining (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected A06240 paraffin-section tonsil caption does not state a fixative (A06240 tissue-IHC caption). That caption reports Tris-EDTA retrieval at pH 9.0 and primary antibody dilution of 1:200, which are starting conditions to validate for lymph node rather than evidence that retrieval is required (A06240 tissue-IHC caption). HPA ICC-IF images show nucleoplasmic localisation, but the supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; lymph-node peroxidase background should be checked with the no-primary control (HPA: Nucleoplasm enhanced; standard chromogenic IHC practice).
HPA tissue IHC evidence for WDHD1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | 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 → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced WDHD1 IHC Tips
Troubleshoot WDHD1 staining in paraffin sections by checking retrieval, nuclear localisation, controls and scoring before interpreting differences between specimens.
How should I retrieve WDHD1 when nuclear staining is weak in paraffin sections?
Use heat-mediated Tris-EDTA at pH 9.0 for antigen retrieval in paraffin sections (datasheet A06240). The selected tonsil image used this retrieval, followed by primary antibody at 1:200 overnight at 4 °C (caption A06240). If staining remains weak, compare a citrate buffer at pH 6.0 on adjacent sections as a fallback, holding antibody concentration and detection conditions constant (standard IHC practice). Judge improvement by nucleoplasmic signal and preservation of tissue morphology, since WDHD1 is nucleoplasmic (UniProt O75717; HPA subcellular). The caption does not report the fixative, so retrieval performance cannot establish fixation sensitivity (caption A06240).
Can fixation explain inconsistent WDHD1 staining across paraffin specimens?
The selected paraffin-embedded tonsil image does not state its fixative, and target-specific WDHD1 fixation sensitivity is unknown (caption A06240). Record each specimen’s fixative, fixation duration and processing history before comparing staining, because fixation can alter epitope accessibility in routine IHC (standard IHC practice). Compare similarly processed sections with the catalog antibody’s pH 9.0 Tris-EDTA retrieval and 1:200 primary dilution as a controlled starting point (datasheet A06240; caption A06240). If one processing group stains poorly, assess tissue morphology and nuclear counterstain alongside the signal before changing retrieval or antibody concentration (standard IHC practice). Do not infer fixation effects from tissue-expression patterns (HPA tissue IHC).
Where should convincing WDHD1 staining appear in chromogenic IHC?
Expect WDHD1 signal in nuclei, particularly the nucleoplasm, when assessing chromogenic sections (UniProt O75717; HPA subcellular). Use the hematoxylin counterstain to distinguish nuclear chromogen from pigment or diffuse cytoplasmic colour (standard IHC practice). HPA describes a ubiquitous nuclear expression profile, with high staining in lymph-node germinal-center cells and appendix glandular cells (HPA tissue IHC). Its tissue entries also report WDHD1 as undetected in adipocytes, so evaluate the named cell population within each specimen (HPA tissue IHC). Because WDHD1 has no annotated transmembrane segment, membrane-only staining conflicts with its recorded localisation and warrants a background-control check (UniProt O75717 topology and localisation; standard IHC practice).
Could an isoform or modified epitope explain discordant WDHD1 IHC staining?
WDHD1 has 2 listed isoforms, but the supplied catalog evidence does not map this antibody’s epitope to either one (UniProt O75717 isoforms; datasheet A06240). Therefore, staining differences alone cannot establish isoform-specific expression; request an epitope map or compare independently validated antibodies if that distinction matters (standard IHC practice). Recorded modifications include phosphoserines at positions 333, 374, 383 and 387, plus acetyllysine at 671 (UniProt O75717 modified residues). Their effect on this antibody’s tissue staining is unknown without epitope information (datasheet A06240; UniProt O75717). First compare matched sections under the documented pH 9.0 retrieval conditions (datasheet A06240).
How can IF help assess an ambiguous chromogenic WDHD1 result?
Use IF as a follow-up localisation check while interpreting the primary chromogenic IHC result, since WDHD1 is reported in the nucleoplasm (UniProt O75717; HPA subcellular). Multiplex WDHD1 with a validated marker for the cell population under study, such as germinal-center cells when examining lymph node (HPA tissue IHC; standard IF practice). Choose a spectrally separated, preferably far-red WDHD1 fluorophore where tissue autofluorescence obscures shorter wavelengths, and inspect single-channel controls (standard IF practice). After fixation, optimise gentle permeabilisation to let antibody reach the nucleoplasmic epitope inside the plasma and nuclear membranes (UniProt O75717 localisation; standard IF practice). Establish IF conditions separately from the catalog’s 1:200 IHC dilution (caption A06240).
What should I check when WDHD1 chromogen covers cytoplasm or tissue edges?
First compare a no-primary control with the WDHD1 section to identify detection-system or endogenous colour unrelated to primary-antibody binding (standard IHC practice). For peroxidase-based chromogenic detection, include an endogenous peroxidase block before applying the detection reagent and inspect the control for residual activity (standard IHC practice). The selected image used primary and secondary antibodies at 1:200, with secondary incubation for 45 minutes at room temperature (caption A06240). If diffuse staining persists, titrate antibody concentration, improve washes and avoid overdeveloping DAB while retaining the same retrieval condition (standard IHC practice; datasheet A06240). WDHD1’s recorded nucleoplasmic location makes broad cytoplasmic colour a reason to recheck specificity (UniProt O75717; HPA subcellular).
How should I score WDHD1 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and count only viable, interpretable nuclei before comparing chromogenic WDHD1 staining (standard IHC practice; UniProt O75717 localisation). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score from 0–300 using intensity categories 0–3 (standard IHC practice). For spatial comparisons, report positive nuclei per mm² of viable compartment and normalise that count to total nuclei or sampled area as appropriate (standard IHC practice). Apply one staining threshold and identical image-acquisition settings across groups, with blinded scoring where feasible (standard IHC practice). Keep cell types separate because HPA reports different staining levels across named populations (HPA tissue IHC).
How do I distinguish true WDHD1 positivity from IHC artefact?
Give greatest weight to nucleoplasmic staining in intact cells, consistent with WDHD1’s recorded location and HPA’s nuclear staining profile (UniProt O75717; HPA tissue IHC and subcellular). Check whether the stained population matches the specimen: HPA reports high staining in lymph-node germinal-center cells but undetected staining in adipocytes (HPA tissue IHC). Exclude edge-heavy staining, necrotic regions and colour present in the no-primary or peroxidase-control section before calling a cell positive (standard IHC practice). Compare against the selected tonsil section’s documented pH 9.0 retrieval and 1:200 primary dilution when troubleshooting (caption A06240). Treat expression comparisons cautiously because HPA reports medium consistency between antibody staining and RNA data (HPA tissue IHC).
Boster reagents
Best WDHD1 / WD repeat and HMG-box DNA-binding protein 1 IHC Antibodies
A06240 has a human tonsil paraffin-section IHC image (catalog image caption); IF is listed without an image (catalog applications). Human and mouse reactivity is listed (catalog reactivity).
Real IHC data
Anti-WDHD1 Antibody
Cat # A06240
A06240 has a human tonsil paraffin-section IHC image and lists IF, with no IF image supplied (catalog image caption; catalog applications; catalog IF images). A06240-1 lists human reactivity and IHC, with no IHC image supplied (catalog reactivity; catalog applications; catalog IHC images).
Which to pick: For tissue IHC, choose A06240 when a documented paraffin-section example matters: its human tonsil image used Tris-EDTA retrieval at pH 9.0 and a 1:200 primary dilution overnight at 4°C; the fixative is unreported (A06240 image caption). For IF, A06240 lists that application at 1:50, but has no IF image; ICC validation is unreported (A06240 catalog applications; catalog IF dilution; catalog IF images). For human and mouse work, A06240 lists both species, while A06240-1 lists human only; both list a rabbit host, and neither reports clonality (catalog reactivity; catalog host; catalog clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75717 (WDHD1_HUMAN, WD repeat and HMG-box DNA-binding protein 1).
- Human Protein Atlas. WDHD1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. WDHD1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. WDHD1 antibody validation summary (2 antibodies).
- Integrative bioinformatics analysis of WDHD1: a potential biomarker for pan-cancer prognosis, diagnosis, and immunotherapy. World journal of surgical oncology 2023 — PMC10523704.
- E2F1-Driven WDHD1 Transcription Enhances Cell-Cycle Progression and Promotes Pancreatic Cancer Progression. Current oncology (Toronto, Ont.) 2026 — PMC13115010.
- WDHD1 is over-expressed in nasopharyngeal carcinoma and may control the expression of ITGAV. FEBS open bio 2023 — PMC9811654.
- WDHD1 is essential for the survival of PTEN-inactive triple-negative breast cancer. Cell death & disease 2020 — PMC7680459.
- PubMed PMID:12508121 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:17081983 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for WDHD1 (WD repeat and HMG-box DNA-binding protein 1). Expected localisation Broad nuclear staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A06240). Validated IHC antibody: A06240 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A06240); 2. Fixation: Image fixative and duration unreported (datasheet A06240); 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: Tris-EDTA pH 9.0 (datasheet A06240); 20 min, 95–100 °C (standard); 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-WDHD1, 1:100-1:300 (datasheet A06240); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt O75717, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A06240, PMC9811654, PMC7680459. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/WDHD1/immunohistochemistry-ihc-guide.