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ANXA10 / Annexin A10 · IHC design guide
Design Immunohistochemistry for ANXA10
Plan ANXA10 IHC on paraffin sections using stomach glandular cells as a strong positive reference and bladder urothelium as a moderate comparator (HPA tissue IHC). This guide covers fixation, antibody dilution, chromogenic detection, controls, and nuclear scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ANXA10 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 stomach and urinary bladder (HPA tissue IHC) | |
| Staining pattern | Stomach glandular and bladder urothelial nuclei (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A13560-1) | |
| Positive control | Stomach+1 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). Selected-image fixative and duration unreported (datasheet A13560-1); verify before use. | |
| Caveat | Bladder staining may be weaker than stomach (HPA tissue IHC) | |
| Regulation | Regulation not annotated (UniProt) | |
| Isoform / epitope | No isoforms or signal/propeptide annotated (UniProt) |
Section 1
Recommended ANXA10 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by four published ANXA10 IHC protocols (PMC7642482; PMC6406441; PMC6686714; PMC6850125).
| Sample | Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A13560-1) |
| Fixation | Image fixative and duration unreported (datasheet A13560-1); 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 A13560-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A13560-1) |
| Primary antibody | Rabbit anti-ANXA10, 2-5 μg/ml (datasheet A13560-1) |
| Primary incubation | Overnight at 4 °C (datasheet A13560-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A13560-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ANXA10-positive staining in glandular cells of stomach (HPA tissue IHC: High). HPA tissue profile: Distinct nuclear expression in stomach and urinary bladder. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet A13560-1). The published protocols provide other retrieval conditions to assess if needed (PMC7642482; PMC6406441; PMC6686714).
Section 2
What Is the Expected ANXA10 Staining Pattern?
In paraffin-section IHC, expect predominantly nuclear ANXA10 staining in stomach glandular cells and urinary bladder urothelial cells (HPA tissue IHC: Enhanced; high in stomach, medium in bladder). HPA describes distinct nuclear expression in these tissues (HPA tissue IHC). UniProt annotates no subcellular location or transmembrane segment, so the nuclear expectation comes from HPA observations, not a UniProt localisation assignment (UniProt Q9UJ72).
| Strong nuclear signal in stomach glandular cells, with surrounding cells comparatively unstained. | This matches the strongest supplied tissue reference: stomach glandular cells are high, and the reported tissue pattern is distinctly nuclear (HPA tissue IHC). Judge signal in the named cells and compartment, rather than treating any brown staining in the section as a positive result. |
| Nuclear signal in urinary bladder urothelial cells that is less intense than the stomach reference. | This is compatible with the supplied medium urothelial staining level (HPA tissue IHC). Medium is a descriptive category, not a calibrated ratio between slides. Compare tissues only after accounting for differences in section and detection conditions (general IHC practice). |
| Predominantly cytoplasmic staining, with no convincing nuclear signal in the expected cells. | This departs from the distinct nuclear tissue pattern (HPA tissue IHC). Consider nonspecific staining or an imaging or scoring artefact; do not assign a validated ANXA10 compartment from tissue IHC alone. HPA reports additional vesicles in ICC-IF, which is a separate assay context (HPA subcellular ICC-IF). |
| Signal chiefly in unrelated cells, such as adipocytes, instead of stomach glandular or bladder urothelial cells. | HPA reports adipocytes as not detected in the listed adipose and breast samples (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity before calling such staining ANXA10; morphology and a detection control can help distinguish those possibilities (general IHC practice). |
| Uniform haze across cells and extracellular spaces, or no signal in the stomach positive reference. | Neither appearance supplies a clear cell-specific nuclear result (HPA tissue IHC: high stomach glandular staining). Broad haze suggests background; an absent positive reference makes a negative test section uninterpretable until staining and detection performance are checked (general IHC practice). |
💡Expected ANXA10 appearanceCall a convincing positive when stomach glandular nuclei show strong staining or bladder urothelial nuclei show medium staining (HPA tissue IHC); diffuse haze or staining confined to unexpected cells is insufficient (general IHC practice).
| Tissue and cell selection | Stomach glandular cells provide the high-staining reference; bladder urothelial cells provide a medium-staining comparison (HPA tissue IHC). Listed not-detected cells can help assess background, but their status does not establish that every cell in an entire organ is negative (HPA tissue IHC). |
| IHC evidence strength | The tissue profile is rated Enhanced for consistency between antibody staining and RNA expression, with external verification pending (HPA tissue IHC). Both listed antibodies have Enhanced IHC status (HPA antibodies). These ratings support the observed pattern without validating every reagent or staining condition. |
| IF/ICC Q: What localisation should an IF experiment assess? | A: HPA reports approved nucleoplasm and nucleoli localisation, plus vesicles as an additional location, in ICC-IF; images are listed for RT-4 and U2OS (HPA subcellular ICC-IF). This informs an IF interpretation but does not make vesicular signal the primary paraffin-section IHC criterion. |
| Topology and molecular annotations | UniProt annotates no transmembrane segment, signal peptide, propeptide, glycosylation sites, or isoforms for the 1–324 ANXA10 chain (UniProt Q9UJ72). Those annotations offer no basis to predict membrane staining, shedding, or an isoform-specific IHC pattern. |
| Retrieval and fixation evidence | Antigen retrieval is a general paraffin-section IHC workflow variable (general IHC practice). The supplied sources give no ANXA10-specific retrieval setting or fixation-sensitivity result; do not infer either from HPA staining levels or UniProt topology (HPA tissue IHC; UniProt Q9UJ72). |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear staining in the stomach positive reference. | A failed staining or detection step is possible; this slide alone cannot establish target absence (general IHC practice; HPA tissue IHC: high stomach glandular staining). | Review section integrity, retrieval and detection controls, then repeat with the antibody's validated IHC-P conditions. Avoid asserting an ANXA10-specific fixation effect, which is unreported in the supplied sources. |
| Bladder signal looks weaker than the stomach signal. | The supplied categories are medium in urothelial cells and high in stomach glandular cells (HPA tissue IHC). | Score urothelial nuclei in their own tissue context before changing conditions. Include the high-staining stomach reference if a run-level positive control is needed (HPA tissue IHC). |
| Only cytoplasmic or membrane-like staining is apparent. | That appearance does not match the reported distinct nuclear tissue pattern (HPA tissue IHC); the cause cannot be assigned from localisation alone. | Check nuclear counterstain alignment, cell morphology and a detection control. Treat vesicles reported by ICC-IF as additional assay-specific evidence, not a substitute for nuclear tissue staining (HPA subcellular ICC-IF). |
| Unexpected cells stain, including adipocytes in a listed negative sample. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA lists those adipocytes as not detected (HPA tissue IHC). | Compare cell identity and staining compartment with the expected positive tissue; assess detection background with a suitable negative reagent control (general IHC practice). |
| Brown signal spreads across the section without cell boundaries. | Diffuse background can arise from detection or blocking conditions (general IHC practice); it is not the distinct nuclear pattern reported for ANXA10 (HPA tissue IHC). | Inspect the negative reagent control and background outside the expected cells; adjust blocking, washes or detection conditions as appropriate for the assay (general IHC practice). |
| A listed not-detected tissue shows scattered positive-looking cells. | The HPA entry describes specific sampled cell types, not a guarantee that every cell in that organ lacks signal (HPA tissue IHC). | Identify the stained cell type and compartment, then compare it with the positive reference and detection control before interpreting it as ANXA10 (general IHC practice). |
Experiment evidence
IHC & IF images showing ANXA10 staining
A13560-1 · IHCIHC analysis of Annexin A10/ANXA10 using anti-Annexin A10/ANXA10 antibody (A13560-1). Annexin A10/ANXA10 was detected in a paraffin-embedded
A13560-1 · IHCIHC analysis of Annexin A10/ANXA10 using anti-Annexin A10/ANXA10 antibody (A13560-1). Annexin A10/ANXA10 was detected in a paraffin-embedded
A13560-1 · IFIF analysis of Annexin A10/ANXA10 using anti-Annexin A10/ANXA10 antibody (A13560-1). Annexin A10/ANXA10 was detected in a paraffin-embedded
PA2077 · IHCAnti-Annexin A10 antibody, PA2077, IHC(P) IHC(P): Human Lung Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109511-ANXA10/tissue/cerebral+cortex#imid_17589835
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000109511-ANXA10/tissue/stomach+1#imid_17589743
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109511-ANXA10/tissue/colon#imid_17589753
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109511-ANXA10/tissue/liver#imid_17589770
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA005469; green = ANXA10, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA005469; green = ANXA10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ANXA10 IHC & IF
🧪Run stomach first: glandular cells should stain strongly (HPA: High in stomach glandular cells). Use adipose tissue as a negative, with adipocytes lacking detectable staining (HPA: Not detected in adipose adipocytes); on the stomach slide, use non-glandular cells to judge background, without assuming they are ANXA10-negative (HPA: stomach result specifies glandular cells).
Positive control tissue: Stomach (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ANXA10 in RT-4, U2OS, with annotated localisation: Nucleoplasm (approved), Nucleoli (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and immunoglobulin-matched rabbit IgG isotype controls (selected-SKU IHC caption: rabbit primary antibody). A validated ANXA10 knockout specimen or peptide competition can test biological specificity; quench endogenous peroxidase for DAB detection and check stomach-section background against the no-primary control (selected-SKU IHC caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected-SKU paraffin-section caption does not state the fixative (selected-SKU IHC caption: fixative not stated). Heat-mediated retrieval in EDTA at pH 8.0 was used in that caption, but whether retrieval is required is unreported (selected-SKU IHC caption: EDTA retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; in stomach sections, assess glandular luminal or mucus-associated background against the controls (HPA: High in stomach glandular cells).
HPA tissue IHC evidence for ANXA10
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Urinary bladder | Urothelial cells | 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 → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ANXA10 IHC Tips
Troubleshoot ANXA10 staining in paraffin sections by checking retrieval, cellular localisation, controls and scoring before changing detection conditions.
Which retrieval conditions should I start with for weak ANXA10 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for this antibody (datasheet A13560-1). The selected paraffin-section image used that buffer, followed by 2 μg/ml primary antibody overnight at 4°C (caption A13560-1). If staining is weak, compare a longer and shorter heating cycle on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Include a stomach section with glandular cells as a positive control, because those cells show high ANXA10 staining (HPA tissue IHC). Check tissue integrity after heating; damaged morphology can make apparent signal gains difficult to interpret (standard IHC practice).
Could fixation explain weak or uneven ANXA10 staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (caption A13560-1). Record the fixative and processing history for each specimen before comparing staining intensity across cases (standard IHC practice). On adjacent sections, keep EDTA at pH 8.0, 2 μg/ml primary antibody and the overnight 4°C incubation constant while assessing the effect of a documented processing difference (datasheet A13560-1; caption A13560-1). Compare preserved glandular morphology and nuclear detail alongside signal, since tissue damage can confound interpretation (standard IHC practice). Do not assign a fixation effect to ANXA10 from its staining pattern alone (standard IHC practice).
Is nuclear ANXA10 staining expected in chromogenic tissue sections?
Yes: tissue IHC shows distinct nuclear expression in stomach and urinary bladder (HPA tissue IHC). ANXA10 is mainly localised to nucleoplasm and nucleoli, with additional vesicular localisation in subcellular imaging (HPA subcellular). Score glandular cells in stomach and urothelial cells in urinary bladder within their respective compartments, rather than treating diffuse staining across all cells as confirmation (HPA tissue IHC; standard IHC practice). A nuclear counterstain helps distinguish genuine nuclear signal from nearby cytoplasmic chromogen (standard IHC practice). If only membrane outlines stain, review morphology, controls and detection background before calling that pattern ANXA10 positive (HPA subcellular; standard IHC practice).
How should I troubleshoot a nuclear-negative result when the antibody epitope is unspecified?
The supplied ANXA10 record lists one 324-aa chain, no annotated isoforms and no transmembrane segment (UniProt Q9UJ72 record). It provides no epitope position, so those annotations cannot establish whether retrieval exposes this antibody’s binding site (UniProt Q9UJ72 record; standard IHC practice). First compare sections retrieved in EDTA at pH 8.0 with a known positive stomach control under the same staining run (datasheet A13560-1; HPA tissue IHC). Keep the documented 2 μg/ml overnight primary incubation fixed while checking retrieval and detection (caption A13560-1). Interpret a failed control as a run-level problem before attributing a negative specimen to epitope variation (standard IHC practice).
How can I check an ANXA10 IHC pattern with multiplex immunofluorescence?
For a separate IF check, pair ANXA10 with a validated marker of stomach glandular cells or urinary bladder urothelial cells, the reported positive populations (HPA tissue IHC; standard IF practice). Choose fluorophores after measuring tissue autofluorescence and reserve a spectrally distinct channel for the cell marker (standard IF practice). ANXA10 has no annotated transmembrane segment, while IF imaging places it mainly in nucleoplasm and nucleoli (UniProt Q9UJ72 record; HPA subcellular). Use permeabilisation appropriate for intracellular nuclear access, then compare with a minimally permeabilised control; the supplied record does not locate this antibody’s epitope (standard IF practice; UniProt Q9UJ72 record). Keep IF observations separate from chromogenic IHC scores (standard assay practice).
What should I change when DAB background obscures ANXA10-positive cells?
Run a no-primary control and inspect it for endogenous peroxidase or detection-related colour before changing the ANXA10 antibody (standard IHC practice). A peroxidase block and controlled DAB development are general chromogenic IHC steps; the selected image used peroxidase-based detection with DAB (standard IHC practice; caption A13560-1). The image also used 10% goat serum blocking, 2 μg/ml primary overnight at 4°C, and secondary incubation for 30 minutes at 37°C (caption A13560-1). If background persists, compare shorter DAB development and stronger washing on adjacent sections (standard IHC practice). Judge improvement against preserved nuclear staining in a positive control (HPA tissue IHC; standard IHC practice).
How should I quantify ANXA10 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before counting: stomach glandular cells and urinary bladder urothelial cells are reported positive, with distinct nuclear tissue staining (HPA tissue IHC). For nuclear chromogenic signal, record the percentage of positive target cells and an H-score of 0–300 using intensity grades 0–3 (standard IHC scoring practice). Normalise each score to evaluable target cells, excluding folds, necrosis and poorly preserved regions, rather than to total section area (standard IHC scoring practice). If counting positive cells per mm², report the analysed viable target-tissue area and sampling rule (standard IHC scoring practice). Use the same threshold and controls across batches (standard IHC practice).
How do I distinguish true ANXA10 positivity from staining artefact?
A convincing result follows the expected cell and compartment pattern: stomach glandular or urinary bladder urothelial cells with nuclear staining (HPA tissue IHC). Mainly nucleoplasmic and nucleolar signal is compatible with subcellular imaging; vesicular signal is also reported, so assess it with cell context and controls (HPA subcellular; standard IHC practice). Treat staining confined to section edges, folds or necrotic regions cautiously, because those patterns can reflect processing artefact (standard IHC practice). Check a no-primary control for endogenous enzyme or detection colour before interpreting diffuse DAB deposits (standard IHC practice). Compare the specimen with positive stomach tissue and a reported negative cell population, such as adipocytes in adipose tissue (HPA tissue IHC).
Boster reagents
Best ANXA10 / Annexin A10 IHC Antibodies
Human paraffin-section IHC images cover appendiceal adenocarcinoma, thyroid cancer, lung adenocarcinoma, placenta and lung cancer (A13560-1 and PA2077 IHC captions); human placenta IF is also shown (A13560-1 IF caption).
Real IHC data
Anti-Annexin A10/ANXA10 Antibody ®
Cat # A13560-1
Real IHC data
Anti-Annexin A10/ANXA10 Antibody ®
Cat # PA2077
A13560-1 has IHC images from human appendiceal adenocarcinoma, thyroid cancer, lung adenocarcinoma and placenta, plus an IF image from human placenta (A13560-1 image captions). PA2077 has a human lung cancer IHC image; its listed IHC reactivity includes human and rat, but the image documents human tissue only (PA2077 IHC caption; catalog applications and reactivity).
Which to pick: Choose A13560-1 for human paraffin-section IHC when its pictured tissues fit your study; its image captions specify EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (A13560-1 IHC captions). For tissue IF, choose A13560-1, which has a human placenta paraffin-section IF image at 5 μg/ml; ICC requires separate validation because the supplied IF image shows tissue (A13560-1 IF caption; catalog applications). Consider PA2077 for rat IHC because rat is listed as reactive, while its IHC image shows human lung cancer only; neither product lists a clone, and neither IHC caption reports the fixative (PA2077 catalog reactivity and IHC caption; A13560-1 and PA2077 catalog clone fields and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UJ72 (ANX10_HUMAN, Annexin A10).
- Human Protein Atlas. ANXA10 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ANXA10 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nucleoli. In addition localized to vesicles..
- Human Protein Atlas. ANXA10 antibody validation summary (2 antibodies).
- Loss of Annexin A10 Expression Is Associated with Poor Prognosis in Early Gastric Cancer. Acta histochemica et cytochemica 2020 — PMC7642482.
- Frequent Alteration of Annexin A9 and A10 in HPV-Negative Head and Neck Squamous Cell Carcinomas: Correlation with the Histopathological Differentiation Grade. Journal of clinical medicine 2019 — PMC6406441.
- Prognostic Significance of Increased Expression of Annexin A10 (ANXA10) in Serous Epithelial Ovarian Cancer. Medical science monitor : international medical journal of experimental and clinical research 2019 — PMC6686714.
- ANXA10 induction by interaction with tumor-associated macrophages promotes the growth of esophageal squamous cell carcinoma. Pathology international 2019 — PMC6850125.
- PubMed PMID:10458909 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ANXA10 (Annexin A10). Expected localisation Nuclear staining in stomach and urinary bladder (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A13560-1). Validated IHC antibody: A13560-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Stomach (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A13560-1); 2. Fixation: Image fixative and duration unreported (datasheet A13560-1); 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 A13560-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A13560-1); 8. Primary antibody: Rabbit anti-ANXA10, 2-5 μg/ml (datasheet A13560-1); 9. Primary incubation: Overnight at 4 °C (datasheet A13560-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A13560-1); verify before use.; Caveat: Bladder staining may be weaker than stomach (HPA tissue IHC); Isoform / epitope: No isoforms or signal/propeptide annotated (UniProt); Regulation: Regulation not annotated (UniProt). Evidence: UniProt Q9UJ72, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm and nucleoli. In addition…, Datasheet A13560-1, PMC7642482, PMC6406441. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANXA10/immunohistochemistry-ihc-guide.