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YWHAH / 14-3-3 protein eta · IHC design guide
Design Immunohistochemistry for YWHAH
Plan YWHAH paraffin IHC around cytoplasmic staining and high signals in Purkinje cells and neuropil (HPA tissue IHC). Keep fixation consistent, and interpret staining cautiously because antibody staining has low concordance with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
YWHAH 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 | Cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining; high in Purkinje cells and neuropil (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04219-2) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Duodenum+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). Selected-image fixative and duration unreported (datasheet A04219-2); verify before use. | |
| Caveat | Antibody staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Mainly expressed in brain (UniProt) | |
| Isoform / epitope | No annotated isoforms; chain spans residues 2–246 (UniProt) |
Section 1
Recommended YWHAH IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A04219-2) is accompanied by one published YWHAH tissue IHC protocol (PMC12549071: IHC methods).
| Sample | Paraffin-embedded mouse cerebellum tissue; fixative not specified (datasheet A04219-2) |
| Fixation | Image fixative and duration unreported (datasheet A04219-2); 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 A04219-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04219-2) |
| Primary antibody | Rabbit anti-YWHAH, 2-5 μg/ml (datasheet A04219-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04219-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04219-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | YWHAH-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a number of tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A04219-2). The published protocol used microwave citrate pH 6.0 with a different anti-YWHAH antibody (PMC12549071: IHC methods).
Section 2
What Is the Expected YWHAH Staining Pattern?
YWHAH should appear mainly cytoplasmic in tissue sections, with strong staining in cerebellar Purkinje cells and cerebral cortex neuropil (HPA tissue IHC: High in both). It has no transmembrane segment (UniProt Q04917 topology). Use these patterns as guides, not absolute pass criteria: HPA rates tissue IHC reliability Uncertain because antibody staining and RNA expression show low consistency (HPA tissue IHC: Uncertain).
| Strong cytoplasmic staining in Purkinje cells; strong signal in cerebral cortex neuropil. | This matches the reported high-signal sites (HPA tissue IHC: High in Purkinje cells and neuropil). Compare stained structures with the counterstain and tissue morphology. HPA’s Uncertain tissue IHC rating means a matching pattern supports interpretation but does not, by itself, establish antibody specificity (HPA tissue IHC: Uncertain). |
| Staining appears only at cell borders or as a dominant nuclear signal in tissue. | Recheck localisation and morphology: the reported tissue pattern is cytoplasmic (HPA tissue IHC: cytoplasmic expression), and YWHAH has no transmembrane segment (UniProt Q04917 topology). A nuclear signal alone is not proof of artefact, since additional nuclear locations are reported in ICC-IF (HPA subcellular: nucleoplasm and nucleoli rim). |
| Strong staining appears in a cell population reported as undetected, while expected positive cells are weak. | Possible explanations include cross-reactivity or endogenous detection activity; check the no-primary control and the location of signal within each cell. For example, HPA reports no detection in lymph node germinal center cells, but that observation is an uncertain IHC reference rather than an absolute negative standard (HPA tissue IHC: Not detected; Uncertain). |
| Colour is diffuse across the section, including spaces outside cells or unrelated structures. | This does not resolve the cytoplasmic pattern reported for YWHAH (HPA tissue IHC: cytoplasmic expression). In general IHC practice, excess primary or secondary reagent, inadequate blocking or washing, and detection-system background can obscure localisation. Compare with a no-primary control before assigning the diffuse colour to YWHAH. |
| Purkinje cells or cerebral cortex neuropil show no detectable stain. | These are useful positive reference structures because HPA reports High staining there (HPA tissue IHC: High in Purkinje cells and neuropil). First check that the intended structures are present, then review the antibody’s IHC-P conditions, retrieval, detection reagents and control results. HPA’s Uncertain rating prevents a negative result from being interpreted alone as target absence (HPA tissue IHC: Uncertain). |
💡Expected YWHAH appearanceA convincing positive shows predominantly cytoplasmic signal, with High staining in cerebellar Purkinje cells or cerebral cortex neuropil (HPA tissue IHC: cytoplasmic; High in both); uniform haze or isolated cell-border staining warrants a background or specificity check (UniProt Q04917 topology).
| Tissue and cell selection | Brain is a useful starting point because UniProt reports expression mainly there, and HPA reports High staining in Purkinje cells and cortex neuropil (UniProt Q04917 tissue specificity; HPA tissue IHC: High). Medium staining in several other tissues and undetected staining in selected cell populations make comparisons cell-specific (HPA tissue IHC: Medium; Not detected). |
| Compartment and topology | Expect the reported cytoplasmic tissue pattern, while interpreting borders against cell morphology (HPA tissue IHC: cytoplasmic expression; UniProt Q04917 topology: no transmembrane segment). UniProt gives no subcellular annotation for YWHAH, so it does not independently validate a precise tissue compartment (UniProt Q04917 subcellular record: not annotated). |
| Strength of IHC evidence | HPA rates tissue IHC Uncertain because antibody staining has low consistency with RNA expression (HPA tissue IHC: Uncertain). The listed antibody CAB025918 also has Uncertain IHC status; HPA061603 has Supported ICC status, which is a separate application assessment (HPA antibodies: CAB025918 IHC Uncertain; HPA061603 ICC Supported). |
| IF/ICC question: can nuclear signal occur? | Yes. HPA reports mainly cytosolic localisation, with additional nucleoplasm and nucleoli-rim signal in ICC-IF (HPA subcellular: cytosol, nucleoplasm, nucleoli rim). Treat that as IF context, not a tissue IHC acceptance rule; HPA cautions that the subcellular summary uses antibodies targeting proteins from multiple genes (HPA subcellular: caution). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive brain structures are blank. | The relevant cells may be absent from the section, or an IHC workflow step may have failed; HPA reports High staining in Purkinje cells and cortex neuropil (HPA tissue IHC: High). | Confirm structures on the counterstain and compare a known-positive section processed in the same run. In general IHC practice, verify the antibody’s stated IHC-P conditions, retrieval, primary incubation and detection reagents before changing interpretation. |
| Signal is weak in a tissue chosen as a positive control. | The sampled cell population may differ from the reported positive population; HPA levels refer to specific cells, not whole organs (HPA tissue IHC: named cells and levels). | Identify the reported cells first. Prefer Purkinje cells or cortex neuropil for a high-signal comparison, and do not expect the same intensity from sites listed as Medium (HPA tissue IHC: High; Medium). |
| Background covers cells and extracellular areas. | General IHC causes include insufficient blocking or washing, concentrated reagents, or endogenous detection activity. A diffuse stain cannot be assigned to the reported cytoplasmic YWHAH pattern (HPA tissue IHC: cytoplasmic expression). | Inspect the no-primary control, then adjust blocking, washes or detection controls according to the assay chemistry. Reassess cell boundaries and intracellular signal after background is reduced. |
| Only nuclei stain in an IHC section. | Tissue IHC is reported as cytoplasmic, although ICC-IF also shows nuclear locations (HPA tissue IHC: cytoplasmic expression; HPA subcellular: additional nucleoplasm and nucleoli rim). | Check the counterstain, no-primary control and positive tissue comparison. Describe reproducible nuclear staining separately; do not score it as the expected tissue pattern solely on the basis of ICC-IF. |
| A reported undetected cell population stains strongly. | Cross-reactivity or endogenous detection activity is possible; HPA’s undetected calls are uncertain tissue IHC observations, not definitive absence (HPA tissue IHC: Not detected; Uncertain). | Confirm the cell identity and compartment, inspect the no-primary control, and compare the result with a reported positive population in the same run. Treat persistent discordance as unresolved specificity evidence. |
| Two tissue sections give conflicting intensity calls. | HPA’s tissue IHC profile has low agreement with RNA expression, and its calls distinguish particular cell populations (HPA tissue IHC: Uncertain; named cells and levels). | Compare the same cell type and compartment in matched staining runs; record section quality and control results. Keep the discrepancy visible in the interpretation instead of treating an HPA level as a fixed threshold. |
Experiment evidence
IHC & IF images showing YWHAH staining
A04219-2 · IHCIHC analysis of YWHAH using anti-YWHAH antibody (A04219-2). YWHAH was detected in a paraffin-embedded section of mouse cerebellum tissue. He
A04219-2 · IHCIHC analysis of YWHAH using anti-YWHAH antibody (A04219-2). YWHAH was detected in a paraffin-embedded section of rat cerebellum tissue. Heat
A04219-2 · IFIF analysis of YWHAH using anti-YWHAH antibody (A04219-2). YWHAH was detected in an immunocytochemical section of U20S cells. Enzyme antigen
A04219 · IFImmunofluorescence analysis of HeLa cells, using 14-3-3 eta Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000128245-YWHAH/tissue/cerebral+cortex#imid_6634440
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000128245-YWHAH/tissue/colon#imid_6633078
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000128245-YWHAH/tissue/liver#imid_6633071
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000128245-YWHAH/tissue/kidney#imid_6633072
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA061603; green = YWHAH, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA061603; green = YWHAH, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for YWHAH IHC & IF
🧪Run cerebellum first; Purkinje cells should stain strongly (HPA: High in cerebellar Purkinje cells). Use duodenum glandular cells as the negative tissue (HPA: Not detected); on the cerebellum slide, treat cells without specific staining as internal background references, since HPA does not identify a defined internal-negative cell type there (HPA: cerebellum row).
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Duodenum (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show YWHAH in A-549, HEK293, U2OS, KOLF2.1J, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (caption: rabbit primary antibody; standard IHC practice). A YWHAH knockout sample, if available, provides a biological negative; quench endogenous peroxidase on cerebellum sections before HRP/DAB detection (standard IHC practice; caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04219-2 cerebellum caption does not state a fixative (selected-SKU caption). The caption uses heat retrieval in EDTA at pH 8.0 on a paraffin section, but does not establish that retrieval is required (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF are easier; assess cerebellar background against the no-primary control when scoring Purkinje cells (HPA: High in Purkinje cells; standard IHC practice).
HPA tissue IHC evidence for YWHAH
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced YWHAH IHC Tips
Troubleshoot YWHAH staining in paraffin sections by comparing retrieval, controls, cell type and compartment before interpreting chromogenic signal.
How should I adjust retrieval when YWHAH staining is weak in paraffin sections?
Start with heat-mediated EDTA at pH 8.0 for paraffin sections stained with A04219-2 (datasheet A04219-2). The selected image documents that buffer but gives no heating duration, so keep the instrument cycle and cooling interval consistent while troubleshooting (datasheet A04219-2; standard IHC practice). Compare each run with a mouse cerebellum reference section processed alongside the samples (datasheet A04219-2). If staining remains weak, test a more stringent retrieval condition on adjacent sections and inspect for tissue loss or diffuse background before adopting it (standard IHC practice).
Could fixation explain weak YWHAH staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative or fixation duration (datasheet A04219-2). Record the fixative, delay before fixation, fixation duration and tissue processing for each specimen, then compare sections handled under similar conditions (standard IHC practice). When staining varies between blocks, run them together with the same retrieval cycle, antibody incubation and detection reagents to identify processing-related variation (standard IHC practice). Do not attribute a weak result to YWHAH-specific fixation sensitivity on the basis of its tissue pattern, topology or modified residues alone (HPA tissue IHC; UniProt Q04917; datasheet A04219-2).
Which staining compartments should I expect for YWHAH in tissue IHC?
Look first for cytoplasmic staining: the tissue profile reports cytoplasmic expression across multiple tissues, while cell imaging places YWHAH mainly in the cytosol (HPA tissue IHC; HPA subcellular). Nuclear signal needs closer validation because cell imaging also reports nucleoplasm and nucleoli-rim localisation, with a caution that the antibodies target proteins from multiple genes (HPA subcellular). Score cytoplasmic and nuclear staining separately in the cell population of interest, using the same counterstain and exposure to chromogen across comparison slides (standard IHC practice). A membrane-only pattern would be unexpected for a protein with no transmembrane segment and warrants review of controls and morphology (UniProt Q04917 topology; standard IHC practice).
How can I investigate an unusual YWHAH staining pattern or suspected epitope issue?
Check the antibody's stated immunogen and species reactivity before interpreting an unusual pattern; the supplied record does not identify the A04219-2 epitope (datasheet A04219-2; UniProt Q04917). YWHAH has no annotated isoforms in this record, so do not assign distinct staining patterns to named splice variants without independent evidence (UniProt Q04917). Its listed modifications include N-acetylglycine at 2 and phosphoserines at 25 and 59, but their effects on this antibody's staining are unknown (UniProt Q04917; datasheet A04219-2). Compare adjacent sections with an independently validated antibody targeting a documented distinct epitope, alongside primary-omission controls, before claiming epitope-dependent localisation (standard IHC practice).
How should I cross-check a YWHAH IHC pattern using IF?
Treat IF as a separate assay requiring its own antibody validation and controls; the selected A04219-2 tissue image documents chromogenic paraffin-section IHC (datasheet A04219-2; standard IF practice). Multiplex YWHAH with a validated marker for the expected cell population, such as Purkinje cells in cerebellum, and check whether signals occupy the same cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores after inspecting unstained tissue for autofluorescence, and include single-label controls to assess bleed-through (standard IF practice). Because YWHAH lacks a transmembrane segment and is mainly cytosolic in cell imaging, use controlled permeabilisation for intracellular access, then compare cytosolic and nuclear patterns cautiously (UniProt Q04917 topology; HPA subcellular; standard IF practice).
What controls help distinguish YWHAH signal from chromogenic background?
Run a primary-omission control through the same secondary antibody, peroxidase detection and DAB development to reveal detection-system background (standard IHC practice). The selected mouse cerebellum image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A04219-2). If diffuse staining persists, compare shorter chromogen development, stronger washing and an appropriate peroxidase block on adjacent sections (standard IHC practice). Examine section edges, folds, necrotic areas and pigment before treating brown deposits as cellular YWHAH staining (standard IHC practice).
How should I quantify YWHAH staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, because the reported tissue signal differs across cell types (HPA tissue IHC; standard IHC practice). For cellular staining, report the percentage of positive cells and an intensity-based H-score, recording cytoplasmic and nuclear scores separately where both are present (standard IHC practice; HPA subcellular). For a tissue-area measure, report positive-cell density per mm² and normalise to the viable, evaluable tissue area rather than the entire image field (standard IHC practice). Keep section thickness, retrieval, imaging settings, chromogen development and positivity threshold consistent, and exclude folds and necrosis using prespecified rules (standard IHC practice).
When is an apparent YWHAH-positive cell likely to be an artefact?
Give greatest weight to staining with clear cellular boundaries in the expected compartment and cell population: cerebellar Purkinje cells have high reported signal, and tissue staining is described as cytoplasmic (HPA tissue IHC). Interpret nuclear staining cautiously because cell imaging reports additional nuclear locations with an antibody-specificity caution (HPA subcellular). An isolated membrane rim, section-edge gradient, necrotic deposit or signal shared by the primary-omission control should prompt review for artefact, including endogenous peroxidase activity (UniProt Q04917 topology; standard IHC practice). The tissue IHC reliability is marked uncertain because antibody staining and RNA expression have low consistency, so corroborate consequential findings independently (HPA tissue IHC).
Boster reagents
Best YWHAH / 14-3-3 protein eta IHC Antibodies
The catalog includes YWHAH antibodies with IHC images from paraffin sections of mouse and rat cerebellum and human stomach, plus IF images from cells and rat cerebellum (catalog image captions).
Real IHC data
Anti-YWHAH Antibody ®
Cat # A04219-2
Real IF data
Anti-14-3-3 eta YWHAH Antibody
Cat # A04219
A04219-2 has IHC images from paraffin sections of mouse and rat cerebellum and human stomach, and IF images from cells and rat cerebellum (A04219-2 image captions). A04219 lists IHC and IF/ICC applications and has a peptide-blocked IF image from HeLa cells (A04219 catalog entry and image caption).
Which to pick: Choose A04219-2 for tissue IHC: its images document paraffin sections with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A04219-2 IHC image captions). Choose A04219 for cell IF/ICC when the peptide-blocked HeLa IF image is useful evidence; its listed IF dilution is 1:200–1:1000 (A04219 catalog entry and image caption). For broader listed species reactivity, A04219-2 includes human, monkey, mouse and rat, although its IHC images document human, mouse and rat only (A04219-2 catalog entry and IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q04917 (1433F_HUMAN, 14-3-3 protein eta).
- Human Protein Atlas. YWHAH tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. YWHAH subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm and nucleoli rim. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. YWHAH antibody validation summary (2 antibodies).
- YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion. International journal of molecular medicine 2025 — PMC12549071.
- Circ_0000144 acts as a miR-1178-3p decoy to promote cell malignancy and angiogenesis by increasing YWHAH expression in papillary thyroid cancer. Journal of otolaryngology - head & neck surgery = Le Journal d'oto-rhino-laryngologie et de chirurgie cervico-faciale 2022 — PMC9330660.
- Ketohexokinase-A acts as a nuclear protein kinase that mediates fructose-induced metastasis in breast cancer. Nature communications 2020 — PMC7595112.
- Caveolin-1 Modulates Mechanotransduction Responses to Substrate Stiffness through Actin-Dependent Control of YAP. Cell reports 2018 — PMC6231326.
- PubMed PMID:8218406 — UniProt-cited evidence.
- PubMed PMID:1578511 — UniProt-cited evidence.
- PubMed PMID:8561965 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for YWHAH (14-3-3 protein eta). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04219-2). Validated IHC antibody: A04219-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Duodenum. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse cerebellum tissue; fixative not specified (datasheet A04219-2); 2. Fixation: Image fixative and duration unreported (datasheet A04219-2); 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 A04219-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04219-2); 8. Primary antibody: Rabbit anti-YWHAH, 2-5 μg/ml (datasheet A04219-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04219-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A04219-2); verify before use.; Caveat: Antibody staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; chain spans residues 2–246 (UniProt); Regulation: Mainly expressed in brain (UniProt). Evidence: UniProt Q04917, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A04219-2, PMC12549071. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/YWHAH/immunohistochemistry-ihc-guide.