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GMNN / Geminin · IHC design guide
Design Immunohistochemistry for GMNN
Use the catalog antibody at 2–5 μg/mL for paraffin-section chromogenic GMNN IHC (datasheet A06060-3). Assess nuclear staining in highly proliferating cells and consider mitotic degradation when comparing fields (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GMNN 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 highly proliferating cells (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in highly proliferating cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06060-3) | |
| 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 (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06060-3) | |
| Caveat | Cell-cycle turnover can alter staining intensity (UniProt) | |
| Regulation | Degraded at metaphase–anaphase (UniProt) | |
| Isoform / epitope | One 1–209 chain; no isoforms annotated (UniProt) |
Section 1
Recommended GMNN IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA retrieval (datasheet A06060-3); the published ACC protocol below uses citrate retrieval (PMC9764478).
| Sample | Paraffin-embedded human gall bladder adenosquamous carcinoma tissue; fixative not specified (datasheet A06060-3) |
| Fixation | Image fixative and duration unreported (datasheet A06060-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A06060-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06060-3) |
| Primary antibody | Rabbit anti-GMNN, 2-5 μg/ml (datasheet A06060-3) |
| Primary incubation | Overnight at 4 °C (datasheet A06060-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06060-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GMNN-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in highly proliferating cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet A06060-3). Citrate at pH 6.0 is the published alternative (PMC9764478).
Section 2
What Is the Expected GMNN Staining Pattern?
GMNN should appear chiefly as nuclear staining in highly proliferating cells on tissue IHC (HPA: tissue profile, reliability Enhanced). High staining is reported in oral mucosal squamous cells, intestinal endocrine cells and testicular pachytene spermatocytes (HPA: tissue IHC). GMNN has no transmembrane segment and can move between cytoplasm and nucleus (UniProt O75496: topology, subcellular location). HPA describes medium consistency between antibody staining and RNA expression (HPA: reliability description).
| Distinct nuclear staining in oral mucosal squamous cells, intestinal endocrine cells or pachytene spermatocytes. | This fits the reported high IHC staining in those cell types and the tissue-level nuclear profile (HPA: tissue IHC). Score the named cells and their nuclei; the reported level does not mean every cell in the section must stain (HPA: cell-specific entries). |
| Predominantly membrane, extracellular or uniformly cytoplasmic chromogen, with little nuclear staining. | A membrane or extracellular pattern conflicts with GMNN's lack of a transmembrane segment and HPA's nuclear tissue profile (UniProt O75496: topology; HPA: tissue IHC). Cytoplasmic staining alone is less decisive because UniProt reports cytoplasmic GMNN; investigate localization before calling it specific (UniProt O75496: subcellular location). |
| Strong staining across cell types that HPA reports as unstained, such as adipocytes or cardiomyocytes. | This conflicts with those cell-specific observations (HPA: adipocytes and cardiomyocytes, not detected). Consider cross-reactivity or endogenous chromogen-generating activity as general IHC possibilities; a positive reaction elsewhere in the same tissue does not establish specificity in these cells (general IHC practice). |
| A diffuse haze covers tissue, obscuring nuclear boundaries and differences between cell types. | The result cannot be matched confidently to the discrete nuclear, cell-specific pattern (HPA: tissue IHC profile). Widespread haze can arise from nonspecific antibody binding or detection background; compare an appropriate negative detection control before interpreting weak nuclei (general IHC practice). |
| No nuclear signal in a section containing an expected positive cell population. | Check that the relevant cells are present: high staining is reported for oral mucosal squamous cells, intestinal endocrine cells and pachytene spermatocytes (HPA: tissue IHC). An absent signal there may indicate assay failure, but the HPA pattern alone cannot identify whether retrieval, antibody incubation or detection caused it (general IHC practice). |
💡Expected GMNN appearanceCall a positive result when high, distinct nuclear chromogen marks the specified oral mucosal squamous cells, intestinal endocrine cells or pachytene spermatocytes (HPA: tissue IHC); broad membrane or extracellular staining is discordant (UniProt O75496: topology; HPA: nuclear profile).
| Compartment and cell-cycle context | HPA reports nuclear expression in highly proliferating tissue cells, while UniProt reports that GMNN can relocate between cytoplasm and nucleus and is degraded during mitosis (HPA: tissue IHC profile; UniProt O75496: location, function). Interpret the cell type and compartment together; do not assume uniform staining across all cells in a section. |
| Cell-specific comparison tissue | HPA reports high staining in several defined cell populations, low staining in esophageal squamous cells, and no detection in adipocytes or cardiomyocytes (HPA: tissue IHC entries). These are comparisons for the named cells, not whole-organ positive or negative labels; they help assess whether the observed distribution is plausible. |
| Antibody validation and assay limits | Three listed antibodies have Enhanced IHC validation, whereas two have no listed IHC status (HPA: antibody validation). The tissue profile has Enhanced reliability but only medium staining-to-RNA consistency (HPA: reliability). Match observations to the validation status of the antibody used; these summaries do not supply a retrieval method, dilution or fixation effect. |
| Topology and other localization | No transmembrane segment or signal peptide is reported (UniProt O75496: topology, processing). HPA ICC-IF supports nucleoplasm as the main location and marks cytosol and basal body as uncertain (HPA: subcellular ICC-IF). The uncertain IF locations should not be treated as established positive compartments for paraffin IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in expected positive nuclei. | The target cells may be absent from the section, or a general IHC step may have failed; HPA supplies a staining pattern, not a cause for assay failure (HPA: tissue IHC; general IHC practice). | Confirm the named positive cells are present, then check the assay's positive control, antigen retrieval, primary-antibody incubation and detection steps (HPA: tissue IHC; general IHC practice). |
| Membrane or extracellular staining dominates. | This distribution conflicts with the reported nuclear tissue pattern and lack of a transmembrane segment (HPA: tissue IHC; UniProt O75496: topology). | Check a negative detection control and examine whether staining follows tissue edges or deposits; treat that pattern as suspect until specific nuclear staining is demonstrated (general IHC practice). |
| Cytoplasmic signal appears without clear nuclear signal. | UniProt allows cytoplasmic GMNN, but HPA's tissue IHC profile is nuclear; HPA ICC-IF calls cytosol uncertain (UniProt O75496: location; HPA: tissue IHC, subcellular ICC-IF). | Review nuclei and named positive cell types first, then compare with a validated positive section before scoring cytoplasm as GMNN in IHC (HPA: tissue IHC; general IHC practice). |
| Many HPA-negative cell types stain. | Staining in adipocytes or cardiomyocytes conflicts with their not-detected entries; cross-reactivity or endogenous detection activity are possible general IHC explanations (HPA: tissue IHC; general IHC practice). | Compare primary-antibody-omission and detection controls, and assess whether the signal remains confined to the reported positive cell types (HPA: tissue IHC; general IHC practice). |
| Diffuse background makes weak nuclei hard to score. | Nonspecific binding or detection background can obscure a nuclear pattern (general IHC practice; HPA: tissue IHC profile). | Check blocking, washes and detection controls; score only nuclei distinguishable from surrounding background and document an uninterpretable section when separation fails (general IHC practice). |
| IF/ICC Q: Should cytosol or basal-body fluorescence count as a positive GMNN result? | HPA supports nucleoplasm as the main ICC-IF location but labels cytosol and basal body uncertain (HPA: subcellular ICC-IF). | Use nucleoplasmic signal as the supported IF/ICC reference; treat cytosol and basal-body signals as tentative and evaluate them with appropriate imaging controls (HPA: subcellular ICC-IF; general IF practice). |
Experiment evidence
IHC & IF images showing GMNN staining
A06060-3 · IHCIHC analysis of Geminin/GMNN using anti-Geminin/GMNN antibody (A06060-3). Geminin/GMNN was detected in a paraffin-embedded section of human
A06060-3 · IHCIHC analysis of Geminin/GMNN using anti-Geminin/GMNN antibody (A06060-3). Geminin/GMNN was detected in a paraffin-embedded section of human
A06060-3 · IFIF analysis of Geminin/GMNN using anti-Geminin/GMNN antibody (A06060-3). Geminin/GMNN was detected in an immunocytochemical section of Caco-
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000112312-GMNN/tissue/cerebral+cortex#imid_13316195
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000112312-GMNN/tissue/duodenum#imid_13316120
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000112312-GMNN/tissue/colon#imid_13316105
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000112312-GMNN/tissue/liver#imid_13316114
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB011458; green = GMNN, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB011458; green = GMNN, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GMNN IHC & IF
🧪Run colon first and look for staining in its endocrine cells (HPA: High in colon endocrine cells). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the colon slide, neighboring cells that remain unstained provide an internal background reference, without assuming every non-endocrine cell is negative.
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GMNN in U-251MG, U2OS, A-431, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit control immunoglobulin appropriate to the primary antibody’s clonality (selected-SKU caption: rabbit primary antibody); confirm specificity with GMNN knockout material or a peptide block if the immunizing peptide is available (standard IHC practice). Block endogenous peroxidase and check for endogenous biotin before using the caption’s biotin-based DAB detection in colon tissue (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not state a fixative (selected-SKU caption: fixative not stated). The paraffin-section example uses heat retrieval in EDTA at pH 8.0, but it does not establish that retrieval is required (selected-SKU caption: EDTA pH 8.0). No supplied comparison shows whether frozen sections or IF are easier; for colon IHC, assess glandular staining against the biotin and peroxidase controls before scoring (selected-SKU caption: paraffin section and biotin-based DAB detection).
HPA tissue IHC evidence for GMNN
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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Rectum | Endocrine cells | High | 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 → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GMNN IHC Tips
Troubleshoot chromogenic GMNN IHC in paraffin sections by checking retrieval, cell compartment, controls and scoring against the supplied tissue evidence.
What retrieval should I try when GMNN staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A06060-3). The selected paraffin-section example used this retrieval before overnight incubation at 4°C with 2 μg/ml catalog antibody, so compare retrieval changes while holding antibody exposure constant (datasheet A06060-3). If staining remains weak, compare a second retrieval condition as a fallback, processing positive-control and test sections together and watching for damaged morphology (standard IHC practice). Record heating and cooling conditions for every run because inconsistent retrieval can change apparent nuclear positivity across sections (standard IHC practice). Evaluate improvement in intact, expected cells rather than accepting stronger diffuse color alone (HPA tissue IHC; standard IHC practice).
How should I troubleshoot fixation-related loss of GMNN staining?
Target-specific GMNN sensitivity to fixation is unknown from the supplied evidence, and the selected paraffin-section caption does not report a fixative (datasheet A06060-3). Document the actual fixative, fixation interval and tissue processing for each specimen before comparing staining intensity across cases (standard IHC practice). Run a consistently processed positive-control section alongside the problem section, then assess whether weak signal coincides with poor morphology or uneven reagent access (standard IHC practice). Adjust processing variables only in controlled comparisons, keeping EDTA retrieval at pH 8.0 and the detection method consistent initially (datasheet A06060-3; standard IHC practice). Do not assign a GMNN-specific fixation effect without a direct comparison (standard IHC practice).
Should GMNN appear in nuclei, cytoplasm, or both?
For paraffin IHC, prioritize interpretable nuclear staining in proliferating cells: the tissue profile describes nuclear expression in highly proliferating cells (HPA tissue IHC). Nucleoplasm is the supported main subcellular location, whereas cytosol and basal body are less certain in the supplied cell-image assessment (HPA subcellular). The protein is also annotated in cytoplasm and nucleus, with potential nuclear relocalization, so cytoplasmic color alone should not automatically be scored as a positive nuclear result (UniProt O75496 localization; standard IHC practice). Compare candidate nuclear signal with counterstained nuclei in intact cells and with matched controls (standard IHC practice). Investigate widespread extracellular or structureless staining as background before assigning biological localization (standard IHC practice).
Could an isoform or masked epitope explain inconsistent staining?
The supplied record annotates 0 isoforms and one 1–209 Geminin chain, so it provides no defined alternate isoform to explain discordant IHC (UniProt O75496). GMNN has no annotated transmembrane segment or glycosylation sites, while several modified residues include acetyllysine and phosphoserines (UniProt O75496). Without a mapped antibody epitope, those modifications cannot be assigned as the cause of a lost or gained stain (UniProt O75496; standard IHC practice). Compare sections with identical EDTA pH 8.0 retrieval and antibody exposure before attributing variation to epitope masking (datasheet A06060-3; standard IHC practice). If discordance persists, document it as unresolved antibody or specimen dependence (standard IHC practice).
How can IF help verify a questionable GMNN IHC pattern?
Use IF/ICC as a separate localization check, pairing GMNN with a validated marker of the expected cell type; the tissue profile includes high staining in endocrine cells and pachytene spermatocytes (HPA tissue IHC; standard IF practice). Choose spectrally distinct fluorophores and inspect unstained tissue for autofluorescence before interpreting apparent overlap (standard IF practice). GMNN has no transmembrane segment and has supported nucleoplasmic localization, so permeabilize appropriately for an intracellular epitope while confirming the antibody’s actual epitope accessibility (UniProt O75496 topology; HPA subcellular; standard IF practice). Include nuclear counterstain and single-channel controls to assess compartment and bleed-through (standard IF practice). Do not transfer the paraffin IHC retrieval conditions into an IF protocol without IF-specific validation (datasheet A06060-3; standard IF practice).
What should I change when the DAB slide stains diffusely?
First compare a no-primary control and an intact positive-control section to separate detection-system color from plausible GMNN staining (standard IHC practice). The selected paraffin example used 10% goat serum block, biotinylated secondary antibody and a streptavidin–biotin system with DAB; preserve those details when reproducing its conditions (datasheet A06060-3). Check peroxidase blocking, endogenous biotin where applicable, washing and DAB development as general chromogenic workflow controls (standard IHC practice). If diffuse color persists, titrate the catalog antibody around the documented 2 μg/ml condition while keeping retrieval and detection constant (datasheet A06060-3; standard IHC practice). Judge improvements by cleaner nuclear signal in intact cells, rather than total brown area (HPA tissue IHC; standard IHC practice).
How should I score GMNN across unevenly stained sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population on the counterstained section before scoring, because the reported tissue pattern emphasizes nuclear expression in highly proliferating cells (HPA tissue IHC; standard IHC practice). Report the percentage of positive eligible nuclei and, when intensity is reproducible, an H-score; for spatial questions, report positive-nucleus density per mm² of viable tissue (standard IHC practice). Normalize counts to the total eligible nuclei or measured viable area, and use the same compartment threshold across cases (standard IHC practice). Exclude necrosis, folds and damaged edges from numerator and denominator before comparing fields (standard IHC practice). Keep retrieval, DAB development and image-analysis thresholds consistent across the comparison set (standard IHC practice).
How can I distinguish genuine GMNN staining from artefact?
A credible chromogenic result places signal in intact nuclei within an appropriate cellular context, matching the reported nuclear expression in highly proliferating cells (HPA tissue IHC; standard IHC practice). Check the compartment carefully: nucleoplasm is the supported main location, although cytoplasmic localization is also annotated, so isolated diffuse cytoplasmic color needs corroboration (HPA subcellular; UniProt O75496 localization; standard IHC practice). Compare candidate positives with a no-primary control and with expected cell populations on the same or a control section (standard IHC practice). Treat edge effects, necrotic deposits and color produced by endogenous enzyme activity as potential artefacts (standard IHC practice). Resolve ambiguous staining through replicated sections and consistent retrieval at pH 8.0 before biological interpretation (datasheet A06060-3; standard IHC practice).
Boster reagents
Best GMNN / Geminin IHC Antibodies
A06060-3 has pictured IHC in human paraffin sections and IF in Caco-2 cells (A06060-3 image captions); the catalog lists Human and Mouse reactivity (catalog: A06060-3).
Real IHC data
Anti-Geminin/GMNN Antibody ®
Cat # A06060-3
A06060-3 has pictured IHC in paraffin sections of human gall bladder adenosquamous carcinoma, lung cancer and rectal cancer, plus IF in Caco-2 cells (A06060-3 image captions). M06060 lists Human reactivity and IHC, ICC and IF applications, but provides no IHC or IF image captions (catalog: M06060).
Which to pick: For tissue IHC, choose A06060-3 when pictured paraffin-section evidence matters; its caption does not report the fixative (A06060-3 IHC captions). For IF/ICC, A06060-3 has a pictured Caco-2 IF example, while M06060 is a monoclonal option listed for ICC/IF without a supplied image (A06060-3 IF caption; catalog: M06060). For cross-species work, A06060-3 lists Human and Mouse reactivity; M06060 lists Human only (catalog: A06060-3, M06060).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75496 (GEMI_HUMAN, Geminin).
- Human Protein Atlas. GMNN tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GMNN subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol and basal body..
- Human Protein Atlas. GMNN antibody validation summary (5 antibodies).
- High expression of GMNN predicts malignant progression and poor prognosis in ACC. European journal of medical research 2022 — PMC9764478.
- Increased FOXJ1 protein expression is associated with improved overall survival in high-grade serous ovarian carcinoma: an Ovarian Tumor Tissue Analysis Consortium Study. British journal of cancer 2023 — PMC9814937.
- Homologous recombination DNA repair deficiency and PARP inhibition activity in primary triple negative breast cancer. Nature communications 2020 — PMC7260192.
- Ex vivo assays to predict enhanced chemosensitization by hyperthermia in urothelial cancer of the bladder. PloS one 2018 — PMC6294360.
- PubMed PMID:9635433 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GMNN (Geminin). Expected localisation Nuclear staining in highly proliferating cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06060-3). Validated IHC antibody: A06060-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human gall bladder adenosquamous carcinoma tissue; fixative not specified…; 2. Fixation: Image fixative and duration unreported (datasheet A06060-3); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A06060-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06060-3); 8. Primary antibody: Rabbit anti-GMNN, 2-5 μg/ml (datasheet A06060-3); 9. Primary incubation: Overnight at 4 °C (datasheet A06060-3). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06060-3); Caveat: Cell-cycle turnover can alter staining intensity (UniProt); Isoform / epitope: One 1–209 chain; no isoforms annotated (UniProt); Regulation: Degraded at metaphase–anaphase (UniProt). Evidence: UniProt O75496, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06060-3, PMC9764478. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GMNN/immunohistochemistry-ihc-guide.