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SHOC2 / Leucine-rich repeat protein SHOC-2 · IHC design guide
Design Immunohistochemistry for SHOC2
Plan chromogenic SHOC2 IHC on paraffin sections using cytoplasmic staining as the tissue reference pattern (HPA tissue IHC). This guide covers control selection, fixation consistency and the catalog antibody A07214-1 starting range of 2–5 μg/mL (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SHOC2 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 tissue staining (HPA tissue IHC); nuclear entry after growth factor stimulation (UniProt) | |
| Staining pattern | Cytoplasmic; some glandular and neuronal cells stain strongly (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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | Growth factors shift nuclear signal (UniProt) | |
| Isoform / epitope | 2 isoforms; no extracellular segment; epitope map unknown (UniProt) |
Section 1
Recommended SHOC2 IHC & IF Protocols
The catalog antibody protocol is accompanied by published SHOC2 IHC methods for breast and pancreatic cancer tissue (PMC6947812; PMC6503676).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07214); 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-SHOC2, 5 μg/mL (datasheet A07214) |
| 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 | SHOC2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a majority of tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the breast study used citrate pH 6.0 (PMC6947812).
Section 2
What Is the Expected SHOC2 Staining Pattern?
SHOC2 should appear mainly in the cytoplasm of many tissue cell types, with nuclear staining also plausible because it can move into the nucleus after growth factor stimulation (HPA: tissue IHC profile; UniProt Q9UQ13: subcellular location). High staining is reported in several glandular, neuronal and squamous epithelial cell populations (HPA: tissue IHC). SHOC2 has no transmembrane segment (UniProt Q9UQ13: topology). HPA rates its tissue staining Approved, with medium consistency against RNA data (HPA: tissue IHC reliability).
| Cytoplasmic chromogenic staining in colon glandular cells or cerebral cortex neurons. | This matches cell populations scored High in those tissues and the predominant cytoplasmic tissue pattern (HPA: tissue IHC). Judge the stained cells and their compartment together; broad tissue expression alone does not make every stained cell a convincing positive (HPA: low RNA tissue specificity; general IHC practice). |
| Nuclear staining accompanies a cytoplasmic signal. | Nuclear localization is biologically plausible: SHOC2 translocates from cytoplasm to nucleus after growth factor stimulation (UniProt Q9UQ13: subcellular location). HPA ICC-IF also places it in the nucleoplasm, but that observation does not establish an expected nuclear intensity for paraffin IHC (HPA: subcellular ICC-IF). |
| A sharp membrane-only rim or extracellular deposit dominates the slide. | Treat this as a compartment mismatch requiring investigation, rather than a confident SHOC2 pattern: the reported locations are cytoplasm and nucleus, and SHOC2 lacks a transmembrane segment (UniProt Q9UQ13: subcellular location and topology). Compare a positive tissue and detection controls before assigning cause (general IHC practice). |
| Strong staining appears in adipocytes or bronchial respiratory epithelial cells. | These cell populations were Not detected in the cited HPA tissue images (HPA: adipose tissue adipocytes; bronchus respiratory epithelial cells). Consider cross-reactivity or endogenous detection activity, while remembering that an HPA nondetection is an observation under its assay conditions, not proof that expression is impossible (general IHC practice; HPA: tissue IHC). |
| Uniform haze covers cells and stroma, or a known-positive tissue has no visible signal. | Haze without cell-specific contrast is background; absence from a High population such as colon glandular cells suggests an assay failure or sample-dependent issue (general IHC practice; HPA: colon glandular cells High). Neither observation alone establishes whether SHOC2 is absent from the specimen (general IHC practice). |
💡Expected SHOC2 appearanceCall the result positive when discrete cytoplasmic staining, potentially with nuclear signal, is visible in an HPA High population such as colon glandular cells; membrane-only rims or uniform tissue-wide haze are suspect (HPA: colon glandular cells High and cytoplasmic tissue profile; UniProt Q9UQ13: nuclear translocation and topology; general IHC practice).
| Cell population and tissue context | HPA scores appendix, colon, duodenum and gallbladder glandular cells, esophageal squamous epithelial cells, and caudate, cerebral cortex and hippocampal neurons High (HPA: tissue IHC). Its low RNA tissue specificity and cytoplasmic staining in a majority of tissues make cell-level interpretation more useful than a simple positive or negative tissue label (HPA: tissue IHC profile and RNA specificity). |
| Growth factor state and compartment | SHOC2 can move from cytoplasm to nucleus after growth factor stimulation (UniProt Q9UQ13: subcellular location). Do not reject a nuclear component solely because the tissue IHC profile emphasizes cytoplasm; do investigate a membrane-only pattern (HPA: tissue IHC profile; UniProt Q9UQ13: topology). |
| Antibody evidence | HPA048134 is Approved for IHC, while HPA009164 is Supported for ICC and has no listed IHC status (HPA: antibody validation). Those labels refer to different applications; ICC support alone does not validate an antibody for paraffin-section interpretation (HPA: antibody validation; general IHC practice). |
| Isoforms and processing | UniProt lists two SHOC2 isoforms, a full-length chain spanning residues 1–582, no signal peptide or propeptide, and no annotated glycosylation sites (UniProt Q9UQ13: isoforms and processing). The supplied evidence gives no epitope map, so it cannot establish whether a given antibody detects both isoforms (UniProt Q9UQ13: isoforms; HPA: antibody validation). |
| IF/ICC Q&A | Q: Can an IF/ICC nuclear signal be expected? A: Yes; HPA reports approved nucleoplasm and supported cytosol localization, and UniProt reports stimulus-dependent nuclear translocation (HPA: subcellular ICC-IF; UniProt Q9UQ13: subcellular location). These observations inform localization, without specifying a paraffin IHC detection condition. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in colon glandular cells or hippocampal neurons. | Both are High in HPA tissue IHC; a blank result may reflect a failed staining run or specimen-specific conditions (HPA: colon glandular cells and hippocampal neurons High; general IHC practice). | Confirm that a concurrently processed positive section stains, then check the primary-antibody step, detection reagents and the assay's validated retrieval settings (general IHC practice). No SHOC2-specific retrieval or fixation sensitivity is supplied. |
| Diffuse brown haze obscures cell boundaries. | Background can arise from nonspecific binding or endogenous chromogenic detection activity (general IHC practice). HPA's cytoplasmic tissue profile does not support interpreting uniform haze as SHOC2 (HPA: tissue IHC profile). | Inspect a no-primary control and the detection system; adjust blocking, washing or detection conditions according to the validated assay (general IHC practice). |
| Signal is restricted to a crisp cell-surface outline. | A membrane-only pattern conflicts with SHOC2's reported cytoplasmic and nuclear locations and lack of a transmembrane segment (UniProt Q9UQ13: subcellular location and topology). | Compare cell-level staining in an HPA High reference tissue and a no-primary control; review nonspecific signal before calling the rim positive (HPA: tissue IHC; general IHC practice). |
| Adipocytes or bronchial respiratory epithelial cells stain strongly. | HPA reports Not detected for those populations under its tissue IHC conditions (HPA: adipose tissue adipocytes; bronchus respiratory epithelial cells). Cross-reactivity or detection-system activity is possible (general IHC practice). | Check whether the signal persists in a no-primary control and whether an HPA High cell population stains with the expected compartment pattern (general IHC practice; HPA: tissue IHC profile). |
| A nuclear component appears alongside cytoplasmic staining. | Nuclear translocation after growth factor stimulation is documented, and HPA ICC-IF reports nucleoplasmic localization (UniProt Q9UQ13: subcellular location; HPA: subcellular ICC-IF). | Score nuclear and cytoplasmic compartments separately and compare the cell-specific pattern with controls; do not classify a nuclear component alone as artefact (general IHC practice; UniProt Q9UQ13: subcellular location). |
| An ICC-supported antibody gives uncertain paraffin IHC staining. | HPA009164 is Supported for ICC but has no listed IHC status; HPA048134 is Approved for IHC (HPA: antibody validation). | Base paraffin IHC interpretation on IHC-specific validation and an appropriate positive section; do not transfer the ICC validation label to IHC (HPA: antibody validation; general IHC practice). |
Experiment evidence
IHC & IF images showing SHOC2 staining
A07214 · IHCImmunohistochemistry of SHOC2 in human spleen tissue with SHOC2 antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108061-SHOC2/tissue/cerebral+cortex#imid_20566291
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108061-SHOC2/tissue/colon#imid_20566233
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108061-SHOC2/tissue/liver#imid_20566234
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108061-SHOC2/tissue/kidney#imid_20566235
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA009164; green = SHOC2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA009164; green = SHOC2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SHOC2 IHC & IF
🧪Run colon first: its glandular cells should stain for SHOC2 (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the colon slide, treat any unstained neighboring cells as an internal background reference, since their SHOC2 status is not specified by the supplied HPA row.
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 SHOC2 in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus SHOC2 knockout material as a biological negative if available (standard IHC practice). Check colon sections for endogenous peroxidase signal during chromogenic detection (standard IHC practice).
⚠️Feasibility: A SHOC2-specific fixation window and antigen-retrieval dependence are unreported in the supplied evidence; the fixative is also unreported in the selected A07214 spleen IHC caption (selected A07214 caption). Frozen sections and IF/ICC are not established as easier than paraffin IHC by the supplied evidence, although ICC-IF images document SHOC2 in the nucleoplasm and cytosol (HPA: subcellular). In colon, inspect luminal mucus and gland edges for nonspecific chromogen deposits (standard IHC practice).
HPA tissue IHC evidence for SHOC2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | 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 → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SHOC2 IHC Tips
Troubleshoot SHOC2 staining in paraffin section IHC by checking retrieval, cellular localisation, controls and scoring before interpreting signal.
How should I retrieve SHOC2 in paraffin sections when staining is weak?
Use Tris-EDTA at pH 9.0 for heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Let sections cool in retrieval buffer, then keep the antibody incubation and chromogenic detection identical across comparison sections (general IHC practice). If staining remains weak, compare a separately processed citrate buffer section at pH 6.0 as a fallback, while checking tissue integrity and nonspecific colour (general IHC practice). Include a positive control with expected staining, such as appendix glandular cells, and assess it alongside the test section (HPA: High in appendix glandular cells).
Could fixation explain variable SHOC2 staining between paraffin sections?
SHOC2-specific sensitivity to fixative or fixation duration is unknown from the supplied evidence (supplied record: no target-specific fixation study). Record the fixative, fixation time and processing history for each section, then compare sections processed alike before attributing a difference to SHOC2 (general IHC practice). The catalog antibody image shows human spleen staining at 5 µg/mL, but its caption does not state the fixative (A07214 caption). If fixation histories differ, repeat staining with the same Tris-EDTA pH 9.0 retrieval and detection conditions, using a shared control section to distinguish run variation from specimen variation (page retrieval rule; general IHC practice).
Should SHOC2 staining appear in cytoplasm, nuclei or cell membranes?
Assess cytoplasmic and nuclear signal separately: SHOC2 is reported in both compartments and can move into the nucleus after growth factor stimulation (UniProt Q9UQ13 subcellular). Tissue IHC shows predominantly cytoplasmic expression across many tissues, while subcellular imaging supports nucleoplasm and cytosol localisation (HPA tissue IHC profile; HPA subcellular). A crisp cell membrane pattern alone deserves scrutiny because SHOC2 has no transmembrane segment (UniProt Q9UQ13 topology). Compare compartments within intact cells, with the same counterstain and detection settings across sections, and avoid assigning a stimulation state from one static stain (general IHC practice; UniProt Q9UQ13 subcellular).
How can I troubleshoot staining when the SHOC2 epitope is unspecified?
SHOC2 has 2 listed isoforms, and the supplied antibody caption does not identify the epitope it recognises (UniProt Q9UQ13 isoforms; A07214 caption). Do not claim that this IHC signal distinguishes isoforms until epitope mapping or isoform-specific validation supports that interpretation (general antibody validation practice). The annotated protein has no signal peptide, transmembrane segment or glycosylation site, so a predicted extracellular staining pattern would need independent confirmation (UniProt Q9UQ13 processing, topology and glycosylation). If staining is unexpected, compare an independently validated SHOC2 antibody or a suitable loss-of-target control under matched retrieval and detection conditions (general IHC practice).
How should I investigate SHOC2 by multiplex IF after reviewing IHC?
For the separate IF/ICC workflow, pair SHOC2 with a marker identifying the cell population being evaluated and include a nuclear counterstain to resolve cytosolic from nucleoplasmic signal (general IF practice; HPA subcellular). Choose fluorophores after inspecting unstained tissue autofluorescence, assigning the weaker expected signal to a cleaner spectral channel where feasible (general IF practice). SHOC2 has no transmembrane segment and is reported in cytosol and nucleoplasm, so test permeabilisation appropriate for intracellular access while monitoring cell morphology (UniProt Q9UQ13 topology; HPA subcellular; general IF practice). Keep single-colour and no-primary controls when judging bleed-through, background and compartment boundaries (general IF practice).
What should I check when SHOC2 chromogenic staining looks diffuse?
Start with a no-primary control and inspect whether colour tracks endogenous peroxidase activity or tissue edges before calling diffuse signal positive (general IHC practice). Confirm that peroxidase blocking, primary antibody concentration, washing and DAB development are consistent across the run (general IHC practice). The selected catalog image reports 5 µg/mL in human spleen, but supplies no fixation or retrieval details for that image (A07214 caption). If background persists, titrate around the documented image concentration, shorten DAB development, and compare intact cellular staining with the no-primary control rather than treating every brown area as SHOC2 (A07214 caption; general IHC practice).
How should I score SHOC2 when cell types and compartments differ? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score cytoplasmic and nuclear staining separately before reviewing outcomes, because both compartments are reported for SHOC2 (UniProt Q9UQ13 subcellular; HPA tissue IHC profile). For cellular staining, report the percentage of positive cells and an H-score using intensity categories 0–3; retain the same thresholds across sections (general IHC practice). If counting spatially distributed positive cells, report density per mm² of evaluable tissue and exclude folds, necrosis and empty areas (general IHC practice). Normalise to the number of evaluable cells or evaluable tissue area within the same defined compartment and cell type, with a shared control across staining runs (general IHC practice).
How do I distinguish plausible SHOC2 signal from staining artefacts?
A plausible result places signal in intact cytoplasm or nuclei, with the compartment and cell type recorded explicitly (UniProt Q9UQ13 subcellular; HPA tissue IHC profile). Compare expected positive populations, such as appendix glandular cells, with a listed undetected population such as adipocytes, while recognising that the HPA tissue IHC assessment has medium consistency with RNA data (HPA: High in appendix glandular cells; HPA: Not detected in adipocytes; HPA reliability description). Treat isolated membrane rims, tissue-edge colour, necrotic areas and no-primary colour as reasons to investigate artefact (UniProt Q9UQ13 topology; general IHC practice). Endogenous peroxidase or excessive DAB development can mimic positivity, so check blocked and no-primary sections before assigning biological meaning (general IHC practice).
Boster reagents
Best SHOC2 / Leucine-rich repeat protein SHOC-2 IHC Antibodies
IHC images show human spleen (A07214 image caption), human liver and liver cancer, and mouse and rat brain (A07214-1 image captions). No IF images are supplied (catalog IF image alts).
Real IHC data
Anti-SHOC2 Antibody
Cat # A07214
A07214 is the sole rendered SKU; its IHC image shows human spleen at 5 μg/mL (A07214 image caption). It is listed for IHC-P and human, mouse and rat reactivity, with IHC validation specified for human samples (A07214 catalog).
Which to pick: For human tissue IHC-P, choose A07214 based on its human spleen image; its caption does not report the fixative (A07214 application list and image caption). For paraffin sections across species, A07214-1 has IHC images of mouse and rat brain and human liver and liver cancer; its captions report EDTA retrieval at pH 8.0 and primary antibody at 2 μg/mL, but do not report the fixative, and neither rabbit antibody has a clone designation supplied (A07214-1 image captions; catalog host and clone fields). Neither SKU has IF/ICC listed or an IF image, so there is no catalog-supported IF/ICC choice (catalog applications and IF image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UQ13 (SHOC2_HUMAN, Leucine-rich repeat protein SHOC-2).
- Human Protein Atlas. SHOC2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SHOC2 subcellular location (ICC-IF): Localized to the nucleoplasm and cytosol..
- Human Protein Atlas. SHOC2 antibody validation summary (2 antibodies).
- SHOC2 is associated with the survival of breast cancer cells and has prognostic value for patients with breast cancer. Molecular medicine reports 2020 — PMC6947812.
- SHOC2 plays an oncogenic or tumor-suppressive role by differentially targeting the MAPK and mTORC1 signals in liver cancer. Life medicine 2024 — PMC11749279.
- The FBXW7-SHOC2-Raptor Axis Controls the Cross-Talks between the RAS-ERK and mTORC1 Signaling Pathways. Cell reports 2019 — PMC6503676.
- PubMed PMID:9674433 — UniProt-cited evidence.
- PubMed PMID:9618511 — UniProt-cited evidence.
- PubMed PMID:10048485 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SHOC2 (Leucine-rich repeat protein SHOC-2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear entry after growth factor stimulation (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A07214 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 A07214); 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-SHOC2, 5 μg/mL (datasheet A07214); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Tissue staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; no extracellular segment; epitope map unknown (UniProt); Regulation: Growth factors shift nuclear signal (UniProt). Evidence: UniProt Q9UQ13, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and cytosol., Datasheet A07214, PMC6947812, PMC6503676. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SHOC2/immunohistochemistry-ihc-guide.