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WSB1 / WD repeat and SOCS box-containing protein 1 · IHC design guide
Design Immunohistochemistry for WSB1
WSB1 shows cytoplasmic tissue staining, including at luminal membranes (HPA tissue IHC). This guide uses colon glandular cells and kidney tubules as positive controls (HPA tissue IHC) and 5 μg/mL catalog antibody as a starting concentration (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
WSB1 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, including luminal membrane (HPA tissue IHC) | |
| Staining pattern | Glandular and tubular cells: cytoplasmic, luminal staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane 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 | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope coverage is unknown (UniProt; datasheet) |
Section 1
Recommended WSB1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with published WSB1 IHC methods for human cancer tissue arrays (PMC4647558) and myocardial paraffin sections (PMC10893653).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07445); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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-WSB1, 5 μg/mL (datasheet A07445) |
| 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 | WSB1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, including luminal membrane. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 at 95–98 °C for 20 min (page retrieval rule). The myocardial protocol reports boiling for 10 min but does not identify the buffer (PMC10893653).
Section 2
What Is the Expected WSB1 Staining Pattern?
In paraffin-section IHC, expect mainly cytoplasmic WSB1 staining, sometimes along the luminal membrane, especially in appendix, colon, gallbladder and prostate glandular cells and kidney tubule cells (HPA: tissue IHC). WSB1 has no transmembrane segment (UniProt Q9Y6I7 topology). The HPA IHC antibody is Approved, but staining has low consistency with RNA expression; treat the pattern as a guide to interpretation, not proof of specificity (HPA: reliability).
| Distinct cytoplasmic staining in glandular cells or kidney tubules, with possible luminal-edge staining. | This matches the reported IHC distribution. Appendix, colon, gallbladder and prostate glandular cells and kidney tubule cells are listed as High; assess the relevant cells rather than assigning one score to the whole section (HPA: tissue IHC). Because HPA reports low consistency between staining and RNA expression, a matching image alone does not establish antibody specificity (HPA: reliability). |
| A predominantly nuclear or other compartment-restricted IHC pattern replaces the expected cytoplasmic pattern. | Flag the result for review: HPA describes tissue IHC as cytoplasmic, including luminal membrane staining (HPA: tissue IHC). Check morphology, counterstain and detection controls before calling it WSB1. HPA separately reports mitochondrial localisation by ICC-IF, so assess IF images against that application-specific observation (HPA: subcellular ICC-IF). |
| Strong staining appears mainly in cells listed as undetected, while the expected cells remain faint. | For example, adipocytes in adipose tissue and alveolar cells in lung are listed as Not detected (HPA: tissue IHC). This mismatch raises possible cross-reactivity or endogenous detection activity; compare a known-positive section and a no-primary control before interpreting the unexpected signal (general IHC practice). |
| Weak colour spreads across many cell types and tissue spaces without a clear cellular boundary. | A broad haze cannot be scored as the HPA cytoplasmic pattern (HPA: tissue IHC). In chromogenic IHC, background may arise from nonspecific antibody binding or endogenous detection activity (general IHC practice). Compare tissue morphology and a no-primary control, then score only discernible cellular staining (general IHC practice). |
| No cellular signal is visible in a section selected for reported high WSB1 staining. | If the relevant glandular or tubule cells are present, this conflicts with the HPA High observations for appendix, colon, gallbladder, kidney and prostate (HPA: tissue IHC). Check that the staining run and positive control worked before reporting absence; HPA's Approved rating and low RNA consistency limit the certainty of any single section (HPA: reliability). |
💡Expected WSB1 appearanceCall a positive IHC result when identifiable glandular cells or kidney tubule cells show clear cytoplasmic staining, possibly at the luminal edge and potentially High in HPA-listed tissues; diffuse haze or dominant staining in listed undetected cells warrants review (HPA: tissue IHC).
| IHC evidence and scoring | The single listed antibody, HPA003293, is IHC Approved; the tissue profile reports low consistency between antibody staining and RNA expression (HPA: antibody validation; HPA: reliability). Use the HPA pattern as a comparison, and interpret intensity in the named cell population (HPA: tissue IHC). |
| Topology and processing | UniProt lists no transmembrane segment, signal peptide, propeptide or glycosylation sites for WSB1 (UniProt Q9Y6I7 topology and processing). The HPA luminal-membrane IHC observation is a staining pattern; it does not establish a membrane-spanning or shed form (HPA: tissue IHC; UniProt Q9Y6I7 topology). |
| Isoforms | UniProt lists 4 isoforms (UniProt Q9Y6I7 isoforms). The supplied evidence does not identify the antibody epitope or establish isoform-specific IHC staining; avoid assigning an IHC pattern to an individual isoform. |
| IF/ICC Q: What location should an IF image be checked against? | A: HPA reports approved mitochondrial localisation in ICC-IF, with images from A-431 and U2OS (HPA: subcellular ICC-IF). This is application-specific evidence; compare the IF image with that localisation while keeping the cytoplasmic tissue-IHC profile as the IHC reference (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-staining reference section shows no signal. | The run may have failed, or the relevant cells may be absent from the examined field (general IHC practice). HPA reports High staining in specified glandular cells and kidney tubules, not every cell in those organs (HPA: tissue IHC). | Confirm the listed cell population on the counterstained section. Review the run control, antibody dilution, antigen-retrieval conditions and chromogen development using the validated IHC procedure (general IHC practice); do not infer WSB1 absence from a failed run. |
| A strong nuclear-only pattern appears. | Its dominant compartment disagrees with the cytoplasmic tissue-IHC profile (HPA: tissue IHC); the supplied evidence does not identify a target-specific cause. | Check tissue morphology and counterstain, compare a reported-positive cell population, and examine a no-primary control for detection signal (general IHC practice). Report the discrepancy if it persists. |
| Listed undetected cell types stain strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice). HPA lists adipocytes in adipose tissue and lung alveolar cells as Not detected (HPA: tissue IHC). | Compare a no-primary control and a reported-positive section in the same run (general IHC practice). Interpret the named cell types separately and avoid calling unexpected staining specific on morphology alone. |
| Diffuse chromogenic background obscures cell boundaries. | Nonspecific binding or endogenous detection activity can obscure cellular staining (general IHC practice); this appearance does not resolve the HPA cytoplasmic pattern (HPA: tissue IHC). | Review blocking, washing, antibody dilution and detection controls using standard IHC practice. Score only signal that remains clearly assigned to intact cells (general IHC practice). |
| Luminal-edge staining is mistaken for proof of a membrane-spanning protein. | HPA includes luminal-membrane staining in its IHC profile, while UniProt lists no transmembrane segment (HPA: tissue IHC; UniProt Q9Y6I7 topology). | Describe the visible edge and cytoplasmic distribution without assigning membrane topology. Compare adjacent cells and the reported glandular pattern before scoring (HPA: tissue IHC). |
| IHC and ICC-IF images seem to show different compartments. | HPA reports cytoplasmic tissue IHC, including luminal membrane, and approved mitochondrial ICC-IF localisation (HPA: tissue IHC; HPA: subcellular ICC-IF). | Evaluate each image against its own HPA application record. Record the method and observed compartment; do not use the ICC-IF result alone to reject an otherwise matching IHC section. |
Experiment evidence
IHC & IF images showing WSB1 staining
A07445 · IHCImmunohistochemistry of WSB1 in human colon tissue with WSB1 antibody at 5 μg/mL.
A07445 · IFImmunofluorescence of WSB1 in human colon tissue with WSB1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109046-WSB1/tissue/cerebral+cortex#imid_1231040
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109046-WSB1/tissue/colon#imid_1230974
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109046-WSB1/tissue/liver#imid_1230975
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109046-WSB1/tissue/kidney#imid_1230968
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003293; green = WSB1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003293; green = WSB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for WSB1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells; selected A07445 tissue-IHC caption: WSB1 staining in human colon). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the colon slide, expect non-glandular areas to show only background relative to glandular cells, without assuming every non-glandular cell is WSB1-negative (HPA: High in colon 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 WSB1 in A-431, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and WSB1-knockout tissue or cells as a biological negative (standard IHC control practice). For chromogenic colon IHC, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window, fixation effect, and antigen-retrieval dependency are unreported in the supplied evidence; the selected A07445 colon IHC caption reports 5 µg/mL but does not state the fixative (selected A07445 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier than paraffin IHC; ICC-IF images are available for A-431 and U2OS cells, with mitochondrial localization reported (HPA: subcellular). In colon, distinguish glandular staining from endogenous peroxidase or detection-related background using the controls above (standard IHC practice).
HPA tissue IHC evidence for WSB1
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 → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Prostate | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced WSB1 IHC Tips
Troubleshoot WSB1 staining in paraffin section IHC by checking retrieval, controls, cellular pattern and scoring before interpreting signal.
How should I retrieve WSB1 antigen when paraffin sections stain weakly?
Use citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). If signal remains weak, confirm that sections were fully deparaffinised, the retrieval bath reached its set temperature, and slides stayed submerged throughout heating (standard IHC practice). Compare an adjusted heating time with the page setting on adjacent sections of the same specimen, while holding antibody dilution and chromogen development constant (standard IHC practice). Use glandular cells in colon as a reference for an expected positive pattern, while recognising that HPA reports low consistency between staining and RNA expression (HPA: High in colon glandular cells; HPA: reliability description).
Could fixation explain weak or uneven WSB1 staining?
WSB1 specific sensitivity to fixation is unknown from the supplied evidence; the colon tissue IHC caption does not state a fixative (A07445 caption: fixative not stated). Record the fixative, time before fixation and fixation duration for each specimen, then compare sections processed under documented conditions (standard IHC practice). For a controlled troubleshooting run, keep section thickness, citrate pH 6.0 retrieval and 20 minute heating constant (page retrieval setting; standard IHC practice). If staining varies with processing, test adjacent sections and judge morphology alongside signal, without assigning the difference to a WSB1 specific fixation effect (standard IHC practice).
Which cellular pattern should I expect in WSB1 IHC?
Assess cytoplasmic staining in the relevant cells, including possible luminal membrane signal in glandular tissue (HPA: tissue IHC profile). Colon glandular cells provide a reported High staining reference, and the A07445 caption documents WSB1 immunohistochemistry in human colon tissue at 5 µg/mL (HPA: colon; A07445 caption). A granular cytoplasmic component may be consistent with the approved mitochondrial location seen by ICC/IF, but that observation does not establish the resolution of chromogenic tissue staining (HPA: subcellular location; standard microscopy practice). Compare cell type and compartment across internal positive and weak areas before calling diffuse nuclear, stromal or edge staining specific (standard IHC practice).
How do I troubleshoot an epitope affected by WSB1 isoforms?
WSB1 has 4 listed isoforms, and its SOCS box spans residues 372–421 (UniProt Q9Y6I7: isoforms and domains). Check the catalog antibody's stated immunogen or mapped epitope against each isoform before interpreting a missing or restricted IHC pattern (standard antibody validation practice). The supplied record reports no signal peptide, no transmembrane segment and no annotated glycosylation sites; it does not specify this antibody's epitope (UniProt Q9Y6I7: processing, topology and glycosylation; supplied antibody evidence). If the epitope is undocumented, compare independently validated reagents or a suitable specificity control under the same retrieval conditions, and report isoform attribution as unresolved (standard IHC practice).
How can IF help investigate an ambiguous WSB1 IHC pattern?
Use IF as a separate check of cellular distribution: HPA reports an approved mitochondrial location in A-431 and U2OS ICC/IF images (HPA: subcellular). For tissue multiplexing, pair WSB1 with a validated marker of the cell type being assessed, such as glandular cells in colon, and inspect channels separately before interpreting overlap (HPA: High in colon glandular cells; standard IF practice). Select a fluorophore channel after measuring tissue autofluorescence in an unstained section, and include single stain controls for bleed through (standard IF practice). Because WSB1 has no annotated transmembrane segment, use permeabilisation appropriate for an intracellular epitope, while checking the antibody's epitope information before assuming its accessibility (UniProt Q9Y6I7: topology; standard IF practice).
What should I change when WSB1 IHC shows widespread background?
First compare a no primary antibody control with the stained section to identify detection reagent or chromogen background (standard IHC practice). Confirm complete deparaffinisation, an appropriate protein block and thorough washes, then shorten chromogen development if staining obscures tissue detail (standard IHC practice). For peroxidase based detection, include a peroxide block and inspect a control for residual endogenous enzyme activity (standard IHC practice). Evaluate whether background follows cut edges, folds or damaged areas rather than the reported cytoplasmic and luminal membrane tissue pattern; colon glandular cells offer a reported High signal reference (standard IHC practice; HPA: tissue IHC profile and colon).
How should I score WSB1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and region before scoring, then use percent positive cells with intensity categories to calculate an H-score where that method fits the study design (standard IHC quantification practice). Alternatively, report positive cell density per mm² of viable tissue when cell counts are more useful than intensity (standard IHC quantification practice). Normalise to the number of eligible cells or viable tissue area, and keep retrieval, antibody conditions, imaging and scoring thresholds identical across specimens (standard IHC quantification practice). Score glandular and nonglandular compartments separately where relevant, since HPA reports High staining in colon glandular cells and low consistency between staining and RNA expression (HPA: colon; HPA: reliability description).
How can I distinguish convincing WSB1 signal from artefact?
Give greater weight to reproducible cytoplasmic signal in an expected cell population, such as colon glandular cells, than to isolated staining at section edges (HPA: tissue IHC profile and colon; standard IHC practice). Compare viable cells with necrotic or damaged regions and inspect no primary and peroxidase block controls when brown deposits are widespread (standard IHC practice). Strong nuclear only staining needs further validation because the supplied tissue profile describes cytoplasmic expression and ICC/IF reports an approved mitochondrial location (HPA: tissue IHC profile; HPA: subcellular). Interpret discordance cautiously: HPA explicitly reports low consistency between antibody staining and RNA expression, so morphology, controls and an independent validation method matter (HPA: reliability description; standard IHC practice).
Boster reagents
Best WSB1 / WD repeat and SOCS box-containing protein 1 IHC Antibodies
The catalog antibody has IHC-P and IF applications in human samples (catalog: IHC-P; IF; Human), with images from human colon tissue (IHC and IF image captions: human colon tissue).
Real IHC data
Anti-WSB1 Antibody
Cat # A07445
A07445 is listed for human IHC-P, with an IHC image of human colon tissue (catalog: Human; IHC-P; IHC image caption: human colon tissue). The same SKU is listed for IF, with an IF image of human colon tissue (catalog: IF; IF image caption: human colon tissue).
Which to pick: For tissue IHC, choose A07445: it is listed for paraffin sections (catalog: IHC-P), and its own IHC image shows human colon tissue at 5 μg/mL (IHC image caption: human colon tissue; 5 μg/mL); the fixative is unreported (IHC image caption). For IF, A07445 has a human colon tissue image at 20 μg/mL (IF image caption: human colon tissue; 20 μg/mL), while ICC validation is unreported (catalog: application list). No cross-species choice is supported: A07445 lists Human reactivity and a Rabbit host, with clonality unreported.
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y6I7 (WSB1_HUMAN, WD repeat and SOCS box-containing protein 1).
- Human Protein Atlas. WSB1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. WSB1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. WSB1 antibody validation summary (1 antibodies).
- WSB1 promotes tumor metastasis by inducing pVHL degradation. Genes & development 2015 — PMC4647558.
- Single-cell protein activity analysis reveals a novel subpopulation of chondrocytes and the corresponding key master regulator proteins associated with anti-senescence and OA progression. Frontiers in immunology 2023 — PMC10077735.
- WSB1, as an E3 ligase, restrains myocardial ischemia-reperfusion injury by activating β-catenin signaling via promoting GSK3β ubiquitination. Molecular medicine (Cambridge, Mass.) 2024 — PMC10893653.
- Ubiqutination via K27 and K29 chains signals aggregation and neuronal protection of LRRK2 by WSB1. Nature communications 2016 — PMC4899630.
- PubMed PMID:10354473 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:10931946 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for WSB1 (WD repeat and SOCS box-containing protein 1). Expected localisation Cytoplasmic, including luminal membrane (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07445 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 A07445); 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 citrate buffer, pH 6.0 , 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-WSB1, 5 μg/mL (datasheet A07445); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Antibody staining has low concordance with RNA expression (HPA tissue IHC); Isoform / epitope: 4 isoforms ; epitope coverage is unknown (UniProt; datasheet); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q9Y6I7, HPA tissue IHC · Approved, HPA ICC-IF · mitochondria., Datasheet A07445, PMC4647558, PMC10893653. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/WSB1/immunohistochemistry-ihc-guide.