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SGO2 / Shugoshin 2 · IHC design guide
Design Immunohistochemistry for SGO2
Plan chromogenic SGO2 IHC in paraffin sections using nuclear staining in testicular pachytene spermatocytes as a benchmark (HPA tissue IHC). Start with the IHC-validated antibody at 1:100–1:300 (datasheet), then score staining by cell type and nuclear location.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SGO2 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 | Nuclear staining in seminiferous ducts (HPA tissue IHC) | |
| Staining pattern | Pachytene spermatocytes: nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Testis+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 A30763) | |
| Caveat | Centromeric signal can disappear at anaphase (UniProt) | |
| Regulation | Centromeric signal lost at anaphase (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage unspecified (UniProt) |
Section 1
Recommended SGO2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is accompanied by four published SGO2 IHC protocols (PMC10414046; PMC12264263; PMC8448842; PMC6554469).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A30763) |
| Fixation | Image fixative and duration unreported (datasheet A30763); 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-SGO2, 1:100 - 1:300 (datasheet A30763) |
| 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 | SGO2-positive staining in pachytene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in seminiferous ducts. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule: nuclear antigen); use each article’s stated conditions when reproducing its staining.
Section 2
What Is the Expected SGO2 Staining Pattern?
SGO2 is a nuclear, centromere and kinetochore protein with no transmembrane segment (UniProt Q562F6 localization and topology). In paraffin section IHC, expect nuclear staining in seminiferous ducts, strongest in pachytene spermatocytes (HPA: tissue profile; High in pachytene spermatocytes). HPA rates its tissue IHC evidence Enhanced, while noting medium consistency between antibody staining and RNA expression; interpret other positive cells with that qualification (HPA: reliability).
| Distinct nuclear staining in pachytene spermatocytes within seminiferous ducts, with relatively little surrounding haze (HPA: nuclear expression in seminiferous ducts; High in pachytene spermatocytes). | This is the clearest tissue level match for SGO2. Nuclear localization agrees with the protein record; centromere or kinetochore localization is phase dependent, so every positive nucleus need not show discrete dots (UniProt Q562F6 localization; HPA: tissue profile). |
| Predominantly cytoplasmic, membranous or extracellular color where nuclear signal is absent (UniProt Q562F6 localization and topology). | That distribution does not match the annotated nuclear and chromosomal compartments or the HPA tissue profile. Check counterstain alignment, chromogen deposits and the detection controls before scoring it as SGO2 (UniProt Q562F6 localization; HPA: tissue profile; general IHC practice). |
| Strong staining in adipocytes or bone marrow hematopoietic cells, especially alongside a weak testis result (HPA: Not detected in adipose adipocytes and bone marrow hematopoietic cells; High in pachytene spermatocytes). | The tissue pattern conflicts with these HPA observations. Consider antibody cross reactivity or endogenous detection activity, then compare a no primary control and the known positive section under matched conditions (HPA: tissue IHC; general IHC practice). |
| Diffuse brown color across nuclei, cytoplasm and section background, without a recognizable cell population (HPA: nuclear tissue profile). | This lacks the cell and compartment selectivity expected from the HPA profile. Review blocking, washes, reagent concentration and section handling; diffuse color alone does not establish SGO2 expression (HPA: tissue profile; general IHC practice). |
| No nuclear signal in pachytene spermatocytes while morphology and counterstain remain assessable (HPA: High in pachytene spermatocytes). | The expected positive control has failed. Check antibody identity and IHC validation, retrieval conditions, primary and detection steps, and a concurrently processed control before calling the specimen negative (HPA: tissue IHC and antibody validation; general IHC practice). |
💡Expected SGO2 appearanceCall a convincing positive when pachytene spermatocyte nuclei stain strongly in testis; isolated strong cytoplasmic or membranous color is suspect (HPA: High in pachytene spermatocytes and nuclear tissue profile; UniProt Q562F6 localization and topology).
| Cell population and cell cycle (HPA: High in pachytene spermatocytes; UniProt Q562F6 localization). | Choose seminiferous ducts containing identifiable pachytene spermatocytes for the positive readout. UniProt describes centromeric localization that changes through division, so absence of obvious puncta in every nucleus is not, by itself, a failed IHC result (HPA: tissue IHC; UniProt Q562F6 localization). |
| Strength of tissue evidence (HPA: Enhanced IHC reliability; medium staining and RNA consistency). | HPA reports Medium staining in appendix and duodenal endocrine cells, fallopian tube non ciliated cells, and rectal enterocytes. These are possible secondary comparisons, but the Enhanced rating still carries the stated RNA consistency limitation (HPA: tissue IHC and reliability). |
| Assay specific localization (HPA: tissue IHC profile and ICC IF subcellular summary). | HPA tissue IHC describes nuclear expression in seminiferous ducts, whereas ICC IF reports mainly nucleoplasm with additional nuclear bodies. Use the latter as context, not as a requirement for resolving nuclear bodies in chromogenic paraffin sections (HPA: tissue IHC; HPA: ICC IF). |
| Antibody validation and isoforms (HPA: antibody validation; UniProt Q562F6 isoforms). | HPA lists Enhanced IHC for HPA035163 and no IHC status for HPA040817; both have Enhanced ICC status. UniProt lists three isoforms, but these records alone do not establish which isoforms an antibody detects in a section (HPA: antibodies; UniProt Q562F6 isoforms). |
| Situation | Likely cause | Next action |
|---|---|---|
| Testis positive control has no nuclear staining (HPA: High in pachytene spermatocytes). | The selected field may lack identifiable pachytene cells, or an IHC reagent step may have failed (HPA: testis cell profile; general IHC practice). | Review morphology in seminiferous ducts, then check the primary antibody, detection reagents and a concurrently processed positive control (HPA: tissue profile; general IHC practice). |
| Signal is weak in a secondary tissue but clear in testis (HPA: High in pachytene spermatocytes; Medium in listed secondary cells). | HPA records lower staining levels in those listed cells; weak staining alone does not imply an assay failure (HPA: tissue IHC). | Score each named cell population separately and retain the testis section as the stronger positive reference (HPA: tissue IHC; general IHC practice). |
| Color is mainly outside nuclei (UniProt Q562F6 localization; HPA: nuclear tissue profile). | Chromogen deposition, background or antibody cross reactivity may mimic signal outside the expected compartment (general IHC practice). | Compare nuclear counterstain and a no primary control; review washes and detection conditions before accepting that pattern (general IHC practice). |
| Negative control cells show strong color (HPA: Not detected in adipose adipocytes and bone marrow hematopoietic cells). | Background binding or endogenous detection activity is possible; HPA's Not detected calls are observations, not an absolute absence of protein (HPA: tissue IHC; general IHC practice). | Check the no primary control and use the detection system's appropriate endogenous activity block; compare matched sections (general IHC practice). |
| All compartments show diffuse staining (HPA: nuclear tissue profile). | Insufficient blocking or washing, or overly concentrated reagents, can reduce IHC contrast (general IHC practice). | Review blocking and washes, and titrate the primary antibody and detection reagents against the positive control (general IHC practice). |
| An ICC IF image shows nuclear bodies, but paraffin IHC shows broader nuclear color (HPA: ICC IF nucleoplasm and nuclear bodies; tissue IHC nuclear profile). | The two HPA assay summaries describe different readouts and do not establish that nuclear bodies must resolve in paraffin IHC (HPA: ICC IF; HPA: tissue IHC). | Judge the paraffin result against the documented nuclear staining and named tissue cell populations; consult the separate IF guide for ICC IF interpretation (HPA: tissue IHC; HPA: ICC IF). |
Experiment evidence
IHC & IF images showing SGO2 staining
A30763 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma, using SGOL2 Antibody. The picture on the right is blocked with th
A08632 · IFIF analysis of SGO2 and Tubulin alpha using anti-SGO2 antibody (A08632) and anti-Tubulin alpha antibody (M03989-3). SGO2 and Tubulin alpha w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163535-SGO2/tissue/cerebral+cortex#imid_8221731
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163535-SGO2/tissue/colon#imid_8220638
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163535-SGO2/tissue/liver#imid_8220631
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163535-SGO2/tissue/kidney#imid_8220640
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035163; green = SGO2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035163; green = SGO2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SGO2 IHC & IF
🧪Run testis first and confirm staining in pachytene spermatocytes (HPA: High in pachytene spermatocytes). Use adipose tissue as the negative tissue (HPA: adipocytes Not detected); on the testis slide, use non-pachytene cells as an internal background reference only where they show no specific staining.
Positive control tissue: Testis (Pachytene spermatocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SGO2 in A-431, U-251MG, U2OS, PC-3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus a peptide-block control using the catalog antibody (A30763 caption: peptide-blocked image). For chromogenic testis IHC, quench endogenous peroxidase and check for background staining in blood-containing areas (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval empirically (standard IHC practice). The selected paraffin-section caption does not state a fixative (A30763 caption: fixative unreported). Frozen sections and IF cannot be judged easier from these sources, and no testis-specific artefact is reported; assess background with the controls above.
HPA tissue IHC evidence for SGO2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Testis | Pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Non-ciliated cells | Medium | Protein (IHC) | HPA → |
| Rectum | Enterocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SGO2 IHC Tips
Use nuclear staining and cell identity to evaluate SGO2 chromogenic IHC, with the catalog image and tissue atlas as separate evidence sources.
Which retrieval conditions should I use when nuclear SGO2 staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). Allow sections to cool in the retrieval buffer before washing, then compare stained nuclei with a no-primary control processed in parallel (standard IHC practice). Score the same cell population across runs because SGO2 is reported in nuclei and at centromeres during defined cell-cycle stages (UniProt Q562F6 localisation). If signal remains weak, test a shorter or longer heating interval on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Record tissue integrity alongside signal so excessive retrieval does not masquerade as improvement (standard IHC practice).
How should I troubleshoot fixation-related loss of SGO2 signal?
Target-specific fixation sensitivity for SGO2 is unknown from the supplied evidence; the catalog image identifies paraffin-embedded human breast carcinoma but does not state its fixative (caption A30763). Start with sections whose fixation and processing history is documented, and compare matched sections when investigating weak nuclear staining (standard IHC practice). Keep retrieval at Tris-EDTA pH 9.0, 95–98 °C for 20 minutes during that comparison (page retrieval setting). Examine morphology and staining together, since damaged or poorly preserved nuclei cannot support reliable nuclear scoring (standard IHC practice). Do not infer a preferred fixative from the tissue atlas staining pattern or SGO2 sequence annotations (HPA tissue IHC; UniProt Q562F6).
What staining pattern supports a credible SGO2 IHC result?
Prioritise nuclear staining: the tissue atlas reports nuclear expression in seminiferous ducts, with high staining in pachytene spermatocytes (HPA tissue IHC). SGO2 is annotated at the nucleus, centromere and kinetochore, with centromeric localisation changing across cell division (UniProt Q562F6 localisation). On chromogenic sections, score the fraction of convincingly stained nuclei within an identified cell population, rather than requiring every nucleus to show a resolved centromeric dot (standard IHC interpretation). Use haematoxylin counterstaining to check nuclear boundaries and cell identity without obscuring weak chromogen (standard IHC practice). Diffuse staining confined to cytoplasm should prompt review of controls and morphology before assignment to SGO2 (UniProt Q562F6 localisation; standard IHC practice).
Can this IHC pattern distinguish SGO2 isoforms or epitope effects?
SGO2 has 3 annotated isoforms, but the supplied catalog caption does not identify which isoform or epitope its antibody detects (UniProt Q562F6 isoforms; caption A30763). The record lists phosphoserine at residue 1144, without evidence that this modification changes staining by the catalog antibody (UniProt Q562F6 modified residues; caption A30763). Treat a peptide-blocked image as support for competition by the supplied peptide, not as proof of isoform specificity (caption A30763; standard IHC interpretation). If staining varies across cell populations, first compare section quality, retrieval and nuclear morphology on adjacent sections (standard IHC practice). Claim isoform-specific IHC only after obtaining epitope mapping and independent isoform validation for the antibody (standard antibody-validation practice).
How can I assess SGO2 by IF alongside the chromogenic IHC result?
For a separate IF/ICC comparison, multiplex SGO2 with a meiotic cell marker such as REC8 and use nuclear morphology to identify the expected spermatocyte population (UniProt Q562F6 REC8 association; HPA tissue IHC). Choose a far-red fluorophore when the specimen has strong shorter-wavelength autofluorescence, and inspect single-channel and no-primary controls before interpreting overlap (standard IF practice). Because SGO2 is nuclear and has no annotated transmembrane segment, optimise post-fixation permeabilisation for access to an intracellular epitope (UniProt Q562F6 localisation and topology; standard IF practice). Document the IF fixative and permeabilisation conditions separately from the paraffin IHC image, whose fixative is unreported (caption A30763). Resolve apparent puncta against a DNA counterstain and the expected cell stage (UniProt Q562F6 localisation; standard IF practice).
How do I separate weak nuclear SGO2 staining from chromogenic background?
Run a no-primary control and inspect tissue edges, folds and damaged regions before calling faint nuclear staining positive (standard IHC practice). Include a peroxidase block before DAB development and check whether residual colour appears in the no-primary section (standard chromogenic IHC practice). Titrate the catalog antibody and shorten chromogen development if broad, uniform colour obscures nuclear boundaries (standard IHC practice). The selected paraffin-section image includes a synthesized-peptide-blocked panel for antibody A30763, which provides a comparison for that exact product (caption A30763). Compare staining in identified cells with the tissue atlas pattern, including high signal in pachytene spermatocytes, while treating its reported consistency with RNA as medium (HPA tissue IHC).
How should I quantify SGO2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring; the tissue atlas reports high SGO2 staining in pachytene spermatocytes and medium staining in several other specified cell types (HPA tissue IHC). For chromogenic IHC, report the percentage of positive nuclei or an H-score derived from nuclear intensity categories, with the threshold set using control sections (standard IHC quantification). If counting discrete positive cells, report density per mm² of evaluable tissue and exclude folds, necrosis and tissue edges (standard IHC quantification). Normalise positive counts to the number of eligible nuclei in each annotated compartment, and apply identical imaging and scoring settings across specimens (standard IHC quantification). Record section and biological sample counts separately to avoid treating fields from one specimen as independent samples (standard IHC quantification).
When should an apparent SGO2-positive result be considered artefactual?
A credible result should fit a nuclear pattern in an appropriate cell population; high staining of pachytene spermatocytes is reported in the tissue atlas (HPA tissue IHC). SGO2 also has centromere and kinetochore annotations, with localisation varying during cell division, so a single snapshot need not show identical nuclear patterns in every cell (UniProt Q562F6 localisation). Reconsider staining dominated by cytoplasm, tissue edges or necrotic areas after checking morphology and no-primary controls (UniProt Q562F6 localisation; standard IHC practice). Colour that persists without primary antibody can indicate endogenous enzyme activity or detection background rather than target staining (standard chromogenic IHC practice). Interpret the peptide-blocked catalog image as product-specific supporting evidence, while keeping its unreported fixative and the atlas’s medium RNA–staining consistency in view (caption A30763; HPA tissue IHC).
Boster reagents
Best SGO2 / Shugoshin 2 IHC Antibodies
The catalog includes anti-SGO2 antibodies with paraffin-section IHC data in human breast carcinoma (A30763 image caption) and IF data in U20S cells (A08632 image caption); listed reactivity spans human, mouse and rat (catalog).
Real IHC data
Anti-SGOL2 SGO2 Antibody
Cat # A30763
Real IF data
Anti-SGO2 Antibody ®
Cat # A08632
A30763 lists IHC and IF for human, mouse and rat, with an IHC image of paraffin-embedded human breast carcinoma (catalog applications, reactivity and A30763 image caption). A08632 lists IF/ICC for human samples, with an IF image of U20S cells (catalog applications, reactivity and A08632 image caption).
Which to pick: Choose A30763 for paraffin-section IHC: it lists IHC at 1:100–1:300 and has a paraffin-section IHC image; the caption does not report the fixative (catalog dilution and A30763 image caption). For IF/ICC, A08632 has a U20S cell IF image at 5 μg/ml, while A30763 also lists IF/ICC (A08632 image caption; catalog applications). For mouse or rat samples, A30763 is the listed polyclonal option with those species in its reactivity list; its displayed IHC image is from a human sample (A30763 dilution_raw, catalog reactivity and A30763 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q562F6 (SGO2_HUMAN, Shugoshin 2).
- Human Protein Atlas. SGO2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SGO2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies..
- Human Protein Atlas. SGO2 antibody validation summary (2 antibodies).
- High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation. Journal of Cancer 2023 — PMC10414046.
- Elevated SGO2 expression in lung adenocarcinoma promotes migration and invasion via MAD2 and correlates with poor prognosis. Scientific reports 2025 — PMC12264263.
- Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas. Scientific reports 2021 — PMC8448842.
- The Ectopic Expression of Meiosis Regulatory Genes in Cutaneous T-Cell Lymphomas (CTCL). Frontiers in oncology 2019 — PMC6554469.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SGO2 (Shugoshin 2). Expected localisation Nuclear staining in seminiferous ducts (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A30763 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Pachytene spermatocytes); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A30763); 2. Fixation: Image fixative and duration unreported (datasheet A30763); 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-SGO2, 1:100 - 1:300 (datasheet A30763); 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 A30763); Caveat: Centromeric signal can disappear at anaphase (UniProt); Isoform / epitope: 3 isoforms; epitope coverage unspecified (UniProt); Regulation: Centromeric signal lost at anaphase (UniProt). Evidence: UniProt Q562F6, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A30763, PMC10414046, PMC12264263. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SGO2/immunohistochemistry-ihc-guide.