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WWC1 / Protein KIBRA · IHC design guide
Design Immunohistochemistry for WWC1
Plan chromogenic WWC1 IHC-P using the catalog antibody's starting concentration of 5 μg/mL (datasheet: A02837). Assess cytoplasmic staining in glandular cells and interpret it with the reported low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
WWC1 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Glandular cells show cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| 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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02837) | |
| Caveat | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression varies by tissue and cell type (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended WWC1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with two published kidney IHC protocols (PMC11896158; PMC11301351).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02837); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| 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-WWC1, 5 μg/mL (datasheet A02837) |
| 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 | WWC1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min when assessing nuclear staining (page retrieval rule; UniProt: nucleus).
Section 2
What Is the Expected WWC1 Staining Pattern?
Expect predominantly cytoplasmic WWC1 staining in glandular cells, especially in appendix, duodenum, endometrium, epididymis, fallopian tube and small intestine (HPA: High in these cells; cytoplasmic expression in most tissues). Perinuclear and nuclear localisation is also reported, so assess compartment alongside tissue and cell identity (UniProt Q8IX03: subcellular location). WWC1 has no transmembrane segment (UniProt Q8IX03: topology). HPA rates tissue IHC Approved but reports low consistency between staining and RNA expression (HPA: reliability).
| Glandular cells show clear cytoplasmic chromogen above nearby background in appendix or duodenum (HPA: High in glandular cells). | This matches the reported IHC pattern when staining follows the glandular cell outlines and controls behave as expected (HPA: cytoplasmic expression in most tissues; general IHC practice). A positive slide supports the run; it does not resolve HPA’s reported staining–RNA inconsistency (HPA: reliability). |
| Staining is exclusively membranous, or outlines nearly every cell while expected glandular cytoplasm is blank. | Treat this as a localisation mismatch and investigate artefact or nonspecific detection (UniProt Q8IX03: no transmembrane segment; HPA: cytoplasmic expression in most tissues; general IHC practice). Do not reject a small perinuclear or nuclear component solely on its location (UniProt Q8IX03: subcellular location). |
| Strong colour appears in adipocytes, cardiomyocytes or bone marrow hematopoietic cells. | These cell populations are reported as Not detected, so prominent staining warrants checks for cross-reactivity or endogenous detection activity (HPA: adipose tissue, heart muscle and bone marrow IHC; general IHC practice). A negative atlas observation is a comparison, not proof that every sample must be blank (HPA: reliability). |
| Chromogen forms a widespread haze across cells, connective tissue and empty slide areas. | A pattern without cellular boundaries cannot establish WWC1 localisation (general IHC practice). Check background with a no-primary control, blocking and wash steps before scoring intensity (general IHC practice). HPA’s cytoplasmic profile describes cellular staining, not diffuse slide background (HPA: tissue IHC profile). |
| A known-positive glandular population has no discernible cytoplasmic signal. | First confirm that the expected cells are present, then review the IHC-P workflow and detection controls (HPA: High in glandular cells of appendix, duodenum, endometrium, epididymis, fallopian tube and small intestine; general IHC practice). Absence in one section alone does not establish true WWC1 loss (general IHC practice). |
💡Expected WWC1 appearanceCall a result positive when identifiable glandular cells show cytoplasmic staining clearly above background, especially in HPA High tissues; isolated strong membrane outlines, staining of reported Not detected cell populations or diffuse haze should prompt review (HPA: tissue IHC profile and listed cell levels; UniProt Q8IX03: topology; general IHC practice).
| Tissue and cell selection | Use a reported High glandular population as a positive reference and score its cells separately from other compartments (HPA: appendix, duodenum, endometrium, epididymis, fallopian tube and small intestine). Breast glandular cells are reported Medium, so a weaker comparison there is plausible (HPA: breast IHC). |
| Evidence strength | The tissue IHC profile is Approved, with low consistency between antibody staining and RNA expression (HPA: reliability). The listed antibodies are IHC Approved; this record does not report IHC Enhanced validation (HPA: HPA038016 and HPA038017 antibody status). Interpret unexpected patterns cautiously and use appropriate controls (general IHC practice). |
| Antigen location and isoforms | WWC1 is a single reported protein chain with 2 isoforms and no transmembrane segment (UniProt Q8IX03: processing, isoforms and topology). The supplied evidence does not locate the catalog antibody’s epitope, so it cannot establish which isoforms it detects or predict an antigen retrieval response (UniProt Q8IX03: supplied record). |
| Endogenous detection activity | Chromogenic IHC can produce signal from tissue activity or nonspecific reagent binding; a no-primary control helps distinguish that background from primary-antibody-dependent staining (general IHC practice). Judge the control in the same cell population being scored (general IHC practice). |
| IF/ICC Q: What location should I expect? | A: HPA reports an approved Golgi apparatus location in ICC-IF images from A-431, U-251MG and U2OS (HPA: subcellular ICC-IF). UniProt also reports cytosol, perinuclear region and nucleus (UniProt Q8IX03: subcellular location). These IF observations do not establish a paraffin-section IHC staining pattern or an IF protocol (HPA: separate tissue IHC and ICC-IF records). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-reference glandular cells remain blank (HPA: High glandular populations). | The expected cells may be absent from the section, or the IHC-P staining or detection run may have failed (general IHC practice). | Confirm the tissue and cell population; check the catalog antibody’s IHC-P instructions, reagent preparation and detection controls before interpreting a biological negative (general IHC practice). |
| Background obscures cytoplasmic boundaries. | Incomplete blocking, washing or detection-system background can mask cellular staining (general IHC practice). | Inspect the no-primary control, then review blocking, wash steps and detection-system controls; score only cellular signal distinguishable from background (general IHC practice). |
| Reported Not detected cells stain as strongly as the positive reference (HPA: listed negative cell populations). | Cross-reactivity or endogenous detection activity is possible; HPA’s low staining–RNA consistency also limits certainty (general IHC practice; HPA: reliability). | Compare the same cell types in a no-primary control and a reported High glandular tissue; investigate persistent primary-dependent staining before assigning WWC1 specificity (HPA: tissue IHC levels; general IHC practice). |
| Signal is predominantly a crisp plasma-membrane rim. | This conflicts with the reported predominantly cytoplasmic IHC profile and WWC1’s lack of a transmembrane segment (HPA: tissue IHC profile; UniProt Q8IX03: topology). | Review counterstain, cell boundaries and controls; document any cytoplasmic component separately and treat isolated rim staining as questionable (general IHC practice). |
| A nuclear or perinuclear component accompanies cytoplasmic staining. | WWC1 has reported nuclear and perinuclear localisation, although HPA describes cytoplasmic expression in most tissues (UniProt Q8IX03: subcellular location; HPA: tissue IHC profile). | Score compartments separately and compare them with controls and the expected cell population; do not classify the entire slide as nonspecific solely because of this component (general IHC practice). |
| IHC shows little Golgi-like concentration despite an ICC-IF report (HPA: Golgi apparatus, ICC-IF). | The HPA Golgi assignment comes from ICC-IF, while its tissue IHC summary describes cytoplasmic staining (HPA: subcellular ICC-IF and tissue IHC). | Interpret the paraffin-section slide against the tissue IHC pattern and its controls; reserve Golgi localisation assessment for the separate IF/ICC design (HPA: tissue IHC and subcellular ICC-IF; general IHC practice). |
Experiment evidence
IHC & IF images showing WWC1 staining
A02837 · IHCImmunohistochemistry of WWC1 in mouse brain tissue with WWC1 antibody at 5 μg/ml.
A02837 · IFImmunofluorescence of WWC1 in mouse brain tissue with WWC1 antibody at 20 μg/ml.
A02837-2 · IFIF analysis of KIBRA/WWC1 using anti-KIBRA/WWC1 antibody (A02837-2). KIBRA/WWC1 was detected in immunocytochemical section of A431 cells. En
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000113645-WWC1/tissue/cerebral+cortex#imid_9742780
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000113645-WWC1/tissue/salivary+gland#imid_9742712
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000113645-WWC1/tissue/colon#imid_9738666
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000113645-WWC1/tissue/liver#imid_9738667
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038016; green = WWC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038016; green = WWC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for WWC1 IHC & IF
🧪Run appendix first: its glandular cells should stain strongly (HPA: High in appendix glandular cells). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the appendix slide, assess non-glandular cells for background-level staining rather than assuming they are WWC1-negative (HPA: appendix result specifies glandular cells).
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 WWC1 in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a concentration-matched isotype control matched to the primary antibody’s host species and clonality, and WWC1-knockout material where available (standard IHC controls). Quench endogenous peroxidase for chromogenic detection and check appendix sections for intrinsic pigment or other background before scoring (standard IHC practice).
⚠️Feasibility: A WWC1-specific fixation window and retrieval dependency are unreported in the supplied evidence; pilot antigen retrieval on matched sections (supplied evidence; standard IHC practice). The selected A02837 mouse-brain IHC caption reports 5 µg/ml but does not report a fixative (selected A02837 caption). ICC-IF images exist for A-431, U-251MG and U2OS, but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC or identify a WWC1-specific tissue artefact (HPA: subcellular record; supplied evidence).
HPA tissue IHC evidence for WWC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced WWC1 IHC Tips
Use compartment, cell type, and matched controls to troubleshoot WWC1 chromogenic staining in paraffin sections (UniProt Q8IX03; HPA tissue IHC).
Which retrieval conditions should I try first for WWC1 paraffin IHC?
Use Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min for WWC1 paraffin IHC (page retrieval setting). Run a known positive section alongside the test section so that a failed retrieval step is distinguishable from low expression (general IHC practice). If staining remains weak, compare citrate pH 6.0 as a fallback while keeping antibody concentration and detection conditions constant (general IHC practice). Inspect morphology after heating, because excessive retrieval can damage sections and complicate compartment scoring (general IHC practice). Record the buffer, temperature, time, and staining outcome for each comparison (general IHC practice).
How should I assess fixation when WWC1 staining is weak?
WWC1-specific fixation sensitivity is unknown from the supplied evidence, so weak staining alone cannot identify fixation as the cause (supplied WWC1 evidence). The mouse brain tissue-IHC caption reports 5 µg/ml antibody but does not state a fixative (selected A02837 caption). For paraffin IHC, compare sections with documented fixation histories while holding retrieval, antibody concentration, and detection constant (general IHC practice). Check tissue morphology and whether weak staining follows fixation batches or occurs only in particular cell populations (general IHC practice). Include a concurrently stained positive section to distinguish a batch-level processing problem from an expected low-signal sample (general IHC practice).
Should I score nuclear, cytoplasmic, or Golgi-like WWC1 staining?
Start by scoring cytoplasmic signal separately from nuclear signal because tissue IHC reports cytoplasmic expression in most tissues, while UniProt also lists nuclear localisation (HPA tissue IHC; UniProt Q8IX03 localisation). UniProt additionally places WWC1 in the perinuclear region, cytosol, and ruffle membrane (UniProt Q8IX03 localisation). A Golgi localisation is reported from subcellular ICC/IF evidence, so treat a compact perinuclear pattern in tissue as a hypothesis for confirmation (HPA subcellular). Assess whether staining occurs in the expected cells rather than scoring every colored structure as positive (HPA tissue IHC; general IHC practice). Document compartment scores separately when patterns differ across samples (general IHC practice).
Could isoforms or epitope accessibility explain discordant WWC1 staining?
WWC1 has 2 listed isoforms, but the supplied evidence does not map this antibody’s epitope to either one (UniProt Q8IX03 isoforms; supplied antibody evidence). Request the immunogen or epitope sequence before interpreting selective tissue staining as isoform-specific (general IHC practice). WWC1 contains WW domains at residues 6–39 and 53–86 and a C2 domain at 658–781 (UniProt Q8IX03 domains). Phosphorylation is annotated at several positions, including 141, 535, and 542, without evidence here that any affects this antibody’s staining (UniProt Q8IX03 modified residues; supplied antibody evidence). Compare retrieval conditions and a positive section before assigning an epitope-based explanation (general IHC practice).
How can I investigate WWC1 localisation by multiplex IF?
Use the separate IF/ICC guide for assay setup, and pair WWC1 with a validated marker for the cell population being examined (general IF practice). For example, neuronal identity matters when examining hippocampus, where tissue IHC reports low neuronal staining (HPA tissue IHC: hippocampus). Choose a fluorophore and detection channel with low tissue autofluorescence, then inspect an unstained section and single-label controls before interpreting overlap (general IF practice). WWC1 lacks a transmembrane segment and is annotated in intracellular compartments, so permeabilise when testing an intracellular epitope, especially nuclear or cytosolic signal (UniProt Q8IX03 topology and localisation; general IF practice). Confirm the antibody’s epitope accessibility under the chosen IF fixation and permeabilisation conditions (general IF practice).
What controls help distinguish WWC1 signal from chromogenic background?
Run a no-primary control to reveal secondary-reagent, endogenous-enzyme, or chromogen background in the same tissue and detection batch (general IHC practice). Include a peroxidase block before a peroxidase-based DAB reaction, and check whether residual color persists in the control (general IHC practice). Reassess blocking and antibody concentration if diffuse staining obscures cell boundaries; the selected mouse brain image reports 5 µg/ml for its own tissue-IHC application (general IHC practice; selected A02837 caption). Adipocytes are reported as not detected by HPA tissue IHC, although that observation does not establish a universal negative control (HPA tissue IHC: adipose tissue). Compare background and candidate positives under identical imaging settings (general IHC practice).
How should I quantify WWC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since WWC1 tissue IHC is predominantly cytoplasmic and UniProt also lists nuclear localisation (HPA tissue IHC; UniProt Q8IX03 localisation). For chromogenic sections, report the percentage of positive target cells and an intensity-based H-score, using the same thresholds across batches (general IHC practice). If spatial distribution is central, report positive-cell density per mm² of viable, annotated tissue (general IHC practice). Normalise counts to the eligible cells or viable tissue area rather than the whole image, and exclude folds, edges, and necrosis consistently (general IHC practice). Keep cytoplasmic and nuclear results separate when both patterns appear (general IHC practice).
When is apparent WWC1 staining credible in a tissue section?
Prioritise reproducible staining within identifiable cells and plausible compartments: HPA describes mostly cytoplasmic tissue staining, while UniProt also lists perinuclear and nuclear localisation (HPA tissue IHC; UniProt Q8IX03 localisation). Compare the stained cell population with tissue context; HPA reports high glandular-cell staining in appendix and low neuronal-cell staining in hippocampus (HPA tissue IHC: appendix and hippocampus). Treat color confined to section edges, folds, necrotic areas, or a no-primary control as possible artefact (general IHC practice). Check endogenous peroxidase when DAB-like color appears without primary antibody (general IHC practice). Interpret weak or absent staining cautiously because HPA reports low consistency between antibody staining and RNA expression data (HPA tissue IHC: reliability description).
Boster reagents
Best WWC1 / Protein KIBRA IHC Antibodies
The catalog shows WWC1 IHC in mouse brain tissue (A02837 image caption) and IF/ICC in A431 cells (A02837-2 image caption); both antibodies list Human, Mouse, and Rat reactivity (catalog reactivity).
Real IHC data
Anti-Protein KIBRA WWC1 Antibody
Cat # A02837
Real IF data
Anti-KIBRA/WWC1 Antibody ®
Cat # A02837-2
A02837 lists IHC-P and IF, with mouse brain tissue images for both applications (catalog applications; image captions). A02837-2 lists IF/ICC and shows staining in A431 cells; it has no listed IHC-P application (catalog applications; IF image caption).
Which to pick: Choose A02837 for paraffin-section tissue IHC: IHC-P is listed, and its own caption shows mouse brain IHC at 5 μg/mL; the fixative is unreported (catalog applications; A02837 IHC image caption). Choose A02837-2 for IF/ICC in cells: its own caption shows A431-cell staining at 5 μg/mL (catalog applications; A02837-2 IF image caption). For work across species, both list Human, Mouse, and Rat reactivity, but the pictured IHC evidence is limited to mouse tissue with A02837; both hosts are Rabbit and neither clone is specified (catalog reactivity, host and clone fields; A02837 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IX03 (KIBRA_HUMAN, Protein KIBRA).
- Human Protein Atlas. WWC1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. WWC1 subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. WWC1 antibody validation summary (2 antibodies).
- Hnrnpk protects against osteoarthritis through targeting WWC1 mRNA and inhibiting Hippo signaling pathway. Molecular therapy : the journal of the American Society of Gene Therapy 2024 — PMC11081807.
- Long non-coding RNA XIST alleviates sepsis-induced acute kidney injury through inhibiting inflammation and cell apoptosis via regulating miR-155-5p/WWC1 axis. The Kaohsiung journal of medical sciences 2022 — PMC11896158.
- WWC1 upregulation accelerates hyperuricemia by reduction in renal uric acid excretion through Hippo signaling pathway. The Journal of biological chemistry 2024 — PMC11301351.
- WWC1 deficiency exacerbates sepsis-induced lung injury by promoting NETosis, M1 and M2b macrophage recruitment, and pyroptosis via YAP1 and STING pathways. Journal of inflammation (London, England) 2026 — PMC13227824.
- PubMed PMID:12559952 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for WWC1 (Protein KIBRA). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A02837 at 5 μg/mL (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: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A02837); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 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-WWC1, 5 μg/mL (datasheet A02837); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02837); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Expression varies by tissue and cell type (HPA tissue IHC). Evidence: UniProt Q8IX03, HPA tissue IHC · Approved, HPA ICC-IF · Golgi apparatus., Datasheet A02837, PMC11896158, PMC11301351. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/WWC1/immunohistochemistry-ihc-guide.