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ANXA2 / Annexin A2 · IHC design guide
Design Immunohistochemistry for ANXA2
Plan chromogenic ANXA2 IHC in paraffin sections using catalog antibody PA1348 (datasheet: PA1348 IHC). Compare membranous and extracellular staining with tissue observations, while accounting for secreted variants that can weaken RNA–protein agreement (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ANXA2 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 | Membranous and extracellular tissue staining (HPA tissue IHC) | |
| Staining pattern | Membranous/extracellular; high in lung macrophages (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PA1348) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adrenal gland+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 | Secreted variants can weaken tissue RNA–protein agreement (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended ANXA2 IHC & IF Protocols
The catalog antibody has a datasheet IHC-P protocol (datasheet: PA1348). The published options below cover parathyroid adenoma, cervical tissue, and endometrium (PMC11279064; PMC10137006; PMC6694070).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet PA1348) |
| Fixation | Image fixative and duration unreported (datasheet PA1348); 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 PA1348); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PA1348) |
| Primary antibody | Rabbit anti-ANXA2, 2-5μg/ml (datasheet PA1348) |
| Primary incubation | Overnight at 4 °C (datasheet PA1348) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PA1348) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ANXA2-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Ubiquitous membranous and extracellular expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: PA1348). If adapting the parathyroid study, assess its microwave pH 6 retrieval separately (PMC11279064).
Section 2
What Is the Expected ANXA2 Staining Pattern?
In paraffin-section IHC, expect ANXA2 mainly at cell borders and extracellular structures (HPA: ubiquitous membranous and extracellular expression). High staining is reported in selected epithelial and glandular cells, cholangiocytes, and lung macrophages (HPA: tissue IHC). ANXA2 has no transmembrane segment; membrane-associated staining does not imply an integral membrane protein (UniProt P07355 topology). HPA rates the tissue pattern Enhanced, while noting that secretion can complicate RNA–protein comparisons (HPA: reliability description).
| Strong staining outlines esophageal squamous cells, bronchial respiratory epithelium, or liver cholangiocytes. | These are supported positive cell populations (HPA: High in esophageal squamous epithelial cells, bronchial respiratory epithelial cells, and liver cholangiocytes). Assess the named cells and their borders, rather than assigning one score to the whole section (general IHC practice). |
| The dominant signal is confined to nuclei, with little membrane or extracellular staining. | A nuclear-only pattern conflicts with the reported tissue profile and approved ICC location (HPA: membranous and extracellular tissue profile; plasma membrane with additional cytosol in ICC). Consider artefact or nonspecific detection; check the positive control and secondary-only control before interpreting it as ANXA2 (general IHC practice). |
| Adrenal glandular cells, cardiomyocytes, or skeletal myocytes stain strongly. | Those cell populations are reported as Not detected (HPA: adrenal glandular cells, heart cardiomyocytes, skeletal muscle myocytes). Check the cell identity and controls; cross-reactivity or endogenous detection activity may explain staining in these cells (general IHC practice). A negative cell population does not make every structure in its tissue negative. |
| Brown signal spreads fairly evenly across cells and empty spaces, obscuring borders. | The expected profile is membranous and extracellular, so extracellular signal alone is not proof of background (HPA: tissue profile; UniProt P07355 subcellular location). A uniform haze that also appears in a secondary-only control suggests nonspecific detection; reassess blocking, washes, and chromogen development (general IHC practice). |
| A correctly identified positive cell population has no visible signal. | Compare with a reported High population, such as lung macrophages or cervical glandular cells (HPA: tissue IHC). A blank result there makes the run difficult to interpret; verify tissue identity, antibody conditions, retrieval, and detection with a run control (general IHC practice). HPA staining is a reference pattern, not a guarantee for every specimen. |
💡Expected ANXA2 appearanceCall the IHC result positive when membrane-associated and/or extracellular signal is clear in a reported High cell population (HPA: tissue IHC profile and levels); dominant nuclear-only staining or strong signal in a reported Not detected cell population warrants a control check (HPA: tissue IHC; general IHC practice).
| Tissue and cell selection | High calls apply to specified cells, including lung macrophages and gallbladder glandular cells (HPA: tissue IHC). Salivary glandular cells are Low, while selected adrenal glandular cells are Not detected (HPA: tissue IHC). Record the cell type as well as the tissue when comparing signal. |
| Membrane and extracellular location | The tissue profile includes both compartments (HPA: tissue IHC). UniProt lists extracellular space, matrix, and basement membrane, as well as movement from cytoplasm to cell surface (UniProt P07355 subcellular location). Interpret extracellular deposits alongside cellular staining and control slides. |
| Antibody validation | Three listed antibodies have Enhanced IHC status: HPA046964, HPA061798, and CAB004311 (HPA: antibody validation). This supports the reported pattern across validation evidence; it does not establish identical staining in every specimen or with an unlisted reagent. |
| RNA and protein comparison | Esophagus is tissue enhanced at RNA level (HPA: RNA specificity). HPA cautions that at least one secreted protein variant can make tissue RNA and protein locations differ (HPA: reliability description). Use observed cell-specific protein staining to judge this IHC section, rather than RNA abundance alone. |
| IF/ICC Q&A: where should signal appear? | Mainly at the plasma membrane, with additional cytosolic signal (HPA: approved ICC-IF locations). This is an ICC-IF location reference, not an IHC intensity standard; the paraffin-section tissue calls come from HPA tissue IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a reported High cell population. | The run may have failed, or the sampled section may lack the identified cells (general IHC practice; HPA: cell-specific High calls). | Confirm cell identity, then review the run control, retrieval, antibody dilution, detection reagents, and development (general IHC practice). |
| Strong nuclear-only signal. | This does not match the reported dominant locations (HPA: tissue membranous/extracellular profile; approved ICC plasma membrane). | Check the positive and secondary-only controls, then reassess detection conditions before scoring it as ANXA2 (general IHC practice). |
| Strong signal in adrenal glandular cells or cardiomyocytes. | These cells are reported as Not detected (HPA: tissue IHC); nonspecific binding or endogenous detection activity is possible (general IHC practice). | Verify morphology and compare secondary-only and detection controls; reassess blocking or detection chemistry if control staining persists (general IHC practice). |
| Diffuse brown haze masks cell borders. | Background from detection or development is possible (general IHC practice); genuine extracellular ANXA2 is also reported (HPA: tissue profile). | Compare with a secondary-only control and inspect whether signal follows anatomical structures; review washes and development (general IHC practice). |
| Only weak signal in salivary glandular cells. | That cell population is reported Low (HPA: salivary glandular cells). | Compare with a reported High cell population on a suitable control section before changing staining conditions (HPA: tissue IHC; general IHC practice). |
| IHC and IF/ICC show different apparent distributions. | The evidence describes tissue IHC as membranous and extracellular, while ICC-IF also reports cytosol (HPA: tissue profile; approved ICC-IF locations). | Evaluate each preparation against its own HPA location reference and its controls; do not transfer an ICC-IF intensity call to paraffin-section IHC (general IHC practice). |
Experiment evidence
IHC & IF images showing ANXA2 staining
PA1348 · IHCIHC analysis of ANXA2 using anti-ANXA2 antibody (PA1348). ANXA2 was detected in a paraffin-embedded section of human liver cancer tissue. He
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000182718-ANXA2/tissue/cerebral+cortex#imid_17071579
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000182718-ANXA2/tissue/lung#imid_17071458
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000182718-ANXA2/tissue/colon#imid_17071505
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000182718-ANXA2/tissue/liver#imid_17071490
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA046964; green = ANXA2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA046964; green = ANXA2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ANXA2 IHC & IF
🧪Run bronchus first and require staining in respiratory epithelial cells (HPA: High in bronchus respiratory epithelial cells). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected in adrenal gland glandular cells); on the bronchus slide, treat neighboring unstained cells as a background reference only, since HPA does not identify a specific internal negative cell type there (HPA: bronchus row).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ANXA2 in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls for the rabbit primary, plus ANXA2-knockout material as a biological negative if available (caption: rabbit anti-ANXA2; standard IHC practice). Block endogenous peroxidase before HRP/DAB detection and check bronchial inflammatory cells for background staining (caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected PA1348 paraffin-section caption does not state the fixative (caption: fixative not stated). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (caption: EDTA pH 8.0). Paraffin-section IHC is directly illustrated; the supplied evidence does not establish frozen sections or IF as easier, and bronchial inflammatory cells warrant attention for endogenous peroxidase background (caption: paraffin-section HRP/DAB; standard IHC practice).
HPA tissue IHC evidence for ANXA2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ANXA2 IHC Tips
Troubleshoot ANXA2 staining in paraffin sections by checking retrieval, cell compartment, controls and scoring before interpreting a chromogenic signal.
What should I change when ANXA2 staining is weak after antigen retrieval?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet PA1348). The PA1348 tissue image used this retrieval before chromogenic detection in a paraffin section of human liver cancer tissue; its caption does not state the fixative (datasheet PA1348). If staining remains weak, compare a small, documented range of heating and cooling conditions on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Include a positive control with an expected cell population, such as respiratory epithelium, and inspect both signal and tissue preservation (HPA: High in bronchial respiratory epithelial cells; standard IHC practice).
Could fixation explain a weak or patchy ANXA2 IHC signal?
Target specific fixation sensitivity is unknown from the supplied evidence: the PA1348 tissue caption describes a paraffin section but does not identify its fixative (datasheet PA1348). Record each specimen’s fixative, fixation interval and processing history, then compare sections processed together under the same EDTA pH 8.0 retrieval conditions (datasheet PA1348; standard IHC practice). Check morphology alongside staining, because poor preservation can make a patchy signal difficult to interpret (standard IHC practice). Use an expected positive cell population and a negative reagent control to distinguish a processing problem from nonspecific chromogen deposition (HPA: High in bronchial respiratory epithelial cells; standard IHC practice).
Where should I expect ANXA2 staining in a tissue section?
Assess membranous and extracellular staining first, while allowing a cytosolic component where cell outlines are clear (HPA: ubiquitous membranous and extracellular expression; HPA subcellular: plasma membrane and cytosol). ANXA2 has no annotated transmembrane segment, and UniProt describes movement from cytoplasm to cell surface through a Golgi independent route (UniProt P07355 topology and subcellular annotation). In the PA1348 paraffin section, compare the signal with morphology rather than assigning every extracellular deposit to a producing cell (datasheet PA1348; standard IHC practice). Record membrane, cytoplasmic and extracellular patterns separately, especially where adjacent cell types or basement membrane could make attribution uncertain (UniProt P07355 subcellular annotation; standard IHC practice).
Can epitope choice explain discordant ANXA2 staining patterns?
The record lists 2 ANXA2 isoforms, but the supplied PA1348 caption does not identify its immunogen or epitope (UniProt P07355 isoforms; datasheet PA1348). Do not infer isoform specificity or recognition of a modified residue from a positive paraffin section alone (standard IHC practice). ANXA2 has annotated phosphorylation at Tyr24 and Ser26, among other modifications, so a defined epitope would matter when comparing reagents or processing conditions (UniProt P07355 modified residues). Request an epitope map or validation data for the antibody before attributing differences between membrane and cytoplasmic staining to isoforms or modification state (HPA subcellular: plasma membrane and cytosol; standard IHC practice).
How should I investigate ANXA2 localisation with IF alongside IHC?
Treat IF as a separate assay and confirm that the antibody works with the chosen fixation and detection conditions before comparing it with PA1348 chromogenic IHC (standard IF practice; datasheet PA1348: tissue IHC caption). Multiplex ANXA2 with a validated marker for the expected cell type, such as a respiratory epithelial marker in bronchus, and check each channel with single stain controls (HPA: High in bronchial respiratory epithelial cells; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence, favoring a channel with clear signal separation (standard IF practice). Because ANXA2 can occur at the cell surface and in cytosol, decide whether to permeabilise according to the mapped antibody epitope and the compartment being tested; that epitope is unreported here (HPA subcellular: plasma membrane and cytosol; datasheet PA1348).
How can I reduce diffuse brown staining without losing ANXA2 signal?
First examine a section lacking primary antibody to locate background from detection reagents, and include a peroxidase block before HRP and DAB development (standard chromogenic IHC practice). The PA1348 image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and an HRP and DAB workflow (datasheet PA1348). If background persists, compare shorter chromogen development or a lower primary concentration on adjacent sections while preserving an expected positive control (standard IHC practice; HPA: High in bronchial respiratory epithelial cells). Evaluate extracellular colour carefully because ANXA2 can occur outside cells, but diffuse staining alone does not establish specific extracellular ANXA2 (HPA: ubiquitous membranous and extracellular expression; standard IHC practice).
How should I score ANXA2 when membrane and extracellular staining coexist? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and an intensity based H score for cellular staining (standard IHC practice; HPA: ubiquitous membranous and extracellular expression). Keep extracellular signal separate as stained area or optical density per mm² of viable, annotated tissue, rather than assigning it to nearby cells (standard image analysis practice). Normalise cellular counts to the number of eligible cells and area measures to the assessed tissue area; exclude folds and necrosis using the same rules across specimens (standard IHC practice). Record retrieval, antibody concentration and development settings because PA1348’s reported 2 μg/ml conditions are part of the comparison context (datasheet PA1348).
What makes an ANXA2 positive result convincing rather than artefactual?
A convincing result follows recognisable cell boundaries or extracellular structures and appears in an expected population, such as bronchial respiratory epithelium or lung macrophages (HPA: ubiquitous membranous and extracellular expression; HPA: High in bronchial respiratory epithelial cells and lung macrophages). Check whether a proposed intracellular pattern agrees with the approved plasma membrane and cytosol localisation (HPA subcellular: plasma membrane and cytosol). Treat staining confined to section edges, folds or necrotic areas cautiously, and use a primary omitted control to investigate endogenous peroxidase or detection background (standard chromogenic IHC practice). Interpret discordance with tissue RNA carefully because a secreted ANXA2 variant can complicate RNA and protein correspondence (HPA: Enhanced reliability description).
Boster reagents
Best ANXA2 / Annexin A2 IHC Antibodies
PA1348 has a human liver cancer paraffin-section IHC image (catalog image caption); M00868 lists IHC and IF/ICC applications with human, mouse, and rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-Annexin A2/ANXA2 Antibody ®
Cat # PA1348
PA1348 is the SKU with a rendered card and its own IHC figure (cards payload). Its image shows chromogenic ANXA2 detection in a paraffin-embedded human liver cancer section (PA1348 image caption).
Which to pick: Choose PA1348 for paraffin-section tissue IHC: its own image documents EDTA retrieval, 2 μg/mL primary antibody, and DAB detection; the fixative is unreported (PA1348 image caption). For IF/ICC, M00868 lists those applications and is rabbit monoclonal, but has no IF image in the payload (M00868 applications/host/clone/image alts). Both list human, mouse, and rat reactivity for cross-species planning; the pictured IHC example is human only (catalog reactivity; PA1348 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P07355 (ANXA2_HUMAN, Annexin A2).
- Human Protein Atlas. ANXA2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ANXA2 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the cytosol..
- Human Protein Atlas. ANXA2 antibody validation summary (3 antibodies).
- Sporadic Parathyroid Adenoma: A Pilot Study of Novel Biomarkers in Females. Medicina (Kaunas, Lithuania) 2024 — PMC11279064.
- Stromal Annexin A2 expression is predictive of decreased survival in pancreatic cancer. Oncotarget 2017 — PMC5739743.
- Protein Profiling in Human Papillomavirus-Associated Cervical Carcinogenesis: Cornulin as a Biomarker for Disease Progression. Current issues in molecular biology 2023 — PMC10137006.
- Expression of annexin A2 in adenomyosis and dysmenorrhea. Archives of gynecology and obstetrics 2019 — PMC6694070.
- PubMed PMID:3013422 — UniProt-cited evidence.
- PubMed PMID:2174397 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ANXA2 (Annexin A2). Expected localisation Membranous and extracellular tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PA1348). Validated IHC antibody: PA1348 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet PA1348); 2. Fixation: Image fixative and duration unreported (datasheet PA1348); 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 PA1348); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PA1348); 8. Primary antibody: Rabbit anti-ANXA2, 2-5μg/ml (datasheet PA1348); 9. Primary incubation: Overnight at 4 °C (datasheet PA1348). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Secreted variants can weaken tissue RNA–protein agreement (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P07355, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet PA1348, PMC11279064, PMC10137006. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANXA2/immunohistochemistry-ihc-guide.