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ANXA2 / Annexin A2 · IHC design guide
Design Immunohistochemistry for ANXA2
Plan ANXA2 staining in paraffin sections using esophageal squamous cells and lung macrophages as positive references (HPA tissue IHC). This guide covers fixation consistency, chromogenic detection, and interpretation of membranous and extracellular staining (HPA tissue IHC), with a catalog antibody IHC range of 2–5 μg/ml (datasheet: PA1348).
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 | Epithelial cells and macrophages; membranous/extracellular (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 paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Secreted variants may separate RNA and protein locations (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | Two isoforms; variant-specific epitope coverage unknown (UniProt) |
Section 1
Recommended ANXA2 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet PA1348). Published IHC protocols cover parathyroid adenoma and endometrium (PMC11279064; 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 EDTA pH 8.0 retrieval for the catalog antibody (datasheet PA1348). The parathyroid protocol provides a microwave pH 6 alternative (PMC11279064 methods).
Section 2
What Is the Expected ANXA2 Staining Pattern?
In paraffin section IHC, expect predominantly membranous and extracellular ANXA2 staining (HPA tissue IHC: ubiquitous membranous and extracellular expression). Strong examples include esophageal squamous epithelium, bronchial respiratory epithelium, liver cholangiocytes and lung macrophages (HPA tissue IHC: High). ANXA2 also occurs at the cell surface, extracellular matrix and basement membrane; it has no transmembrane segment (UniProt P07355: subcellular location and topology). HPA rates the tissue staining evidence Enhanced (HPA tissue IHC: reliability).
| Strong cell borders or extracellular staining in esophageal squamous cells, with interpretable counterstain. | This fits a known high staining cell population and the reported membranous and extracellular profile (HPA tissue IHC: Esophagus High; profile). Compare neighboring cells before calling the entire section uniformly positive (standard IHC interpretation). |
| Predominantly nuclear staining, with little membranous or extracellular signal. | This is discordant with the reported tissue profile and approved IF locations; investigate nonspecific detection before scoring it as ANXA2 (HPA tissue IHC: profile; HPA ICC-IF: plasma membrane and cytosol). |
| Strong staining in adrenal glandular cells while an on-slide known-positive population stains as expected. | HPA reports adrenal glandular cells as Not detected; assess antibody cross-reactivity or endogenous chromogen-generating activity (HPA tissue IHC: adrenal glandular cells Not detected; standard IHC practice). |
| Diffuse chromogen covers tissue and blank regions, obscuring cell borders. | The distribution cannot be scored against ANXA2's reported membranous and extracellular pattern (HPA tissue IHC: profile). Check blocking, washes and detection controls for nonspecific background (standard IHC practice). |
| No signal in esophageal squamous epithelium or bronchial respiratory epithelium. | Both are reported High, so first suspect an assay failure, while recognizing that individual sections and antibody performance require controls (HPA tissue IHC: Esophagus and Bronchus High; standard IHC practice). |
💡Expected ANXA2 appearanceCall positive when defined membranous or extracellular staining is clear in an HPA High cell population, such as esophageal squamous epithelium; diffuse nuclear-only or field-wide chromogen fails that pattern check (HPA tissue IHC: profile and Esophagus High; HPA ICC-IF: approved locations; standard IHC interpretation).
| Compartment and topology | ANXA2 has no transmembrane segment despite cell surface localization; a membrane-associated outline is plausible without implying integral membrane insertion (UniProt P07355: topology and subcellular location). Extracellular matrix and basement membrane localization can also contribute staining (UniProt P07355: subcellular location). |
| Cell-specific reference levels | HPA reports High staining in liver cholangiocytes and lung macrophages, Low staining in salivary glandular cells, and Not detected in skeletal myocytes (HPA tissue IHC: cell-specific levels). Judge the annotated cell population, not the organ name alone (standard IHC interpretation). |
| Protein versus RNA | HPA calls RNA tissue enhanced in esophagus but protein staining ubiquitous, and cautions that a secreted variant complicates RNA–protein correlation (HPA tissue IHC: RNA specificity, profile and reliability description). RNA abundance alone should not determine IHC scoring (standard IHC interpretation). |
| Antibody evidence | Three listed antibodies have Enhanced IHC status; this supports the reported tissue pattern but does not validate every reagent or staining condition (HPA antibodies: HPA046964, HPA061798 and CAB004311 IHC Enhanced). Check the antibody actually used against its own controls (standard IHC practice). |
| Isoforms and processing | UniProt lists two isoforms and an ANXA2 chain spanning residues 2–339, but the payload gives no antibody epitope (UniProt P07355: isoforms and processing). Do not assign compartment differences to isoform recognition or processing from this evidence alone. |
| IF/ICC Q&A: Where should ANXA2 appear? | Mainly at the plasma membrane, with additional cytosolic signal in the HPA IF record (HPA ICC-IF: approved locations). This answers the localization question only; paraffin section IHC staining is described by the tissue profile (HPA tissue IHC: profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive esophageal epithelium has no visible chromogen. | A failed staining or detection step is plausible when an HPA High population is blank (HPA tissue IHC: Esophagus High; standard IHC practice). | Review the catalog antibody's IHC-P instructions, reagent controls and chromogen development; repeat with a known-positive section (standard IHC practice). |
| An adrenal glandular cell population stains strongly. | That population is reported Not detected; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: adrenal glandular cells Not detected; standard IHC practice). | Compare a no-primary control and the expected positive population; address endogenous detection activity if the control also stains (standard IHC practice). |
| Most nuclei stain while membrane outlines remain faint. | Nuclear dominance conflicts with reported membranous tissue expression and approved plasma membrane IF localization (HPA tissue IHC: profile; HPA ICC-IF: main location). | Check primary antibody specificity and no-primary background, then score only staining with a supported cellular distribution (standard IHC practice). |
| Extracellular color is widespread and hard to assign to cells. | Extracellular ANXA2 is plausible, but diffuse background can obscure its distribution (UniProt P07355: extracellular matrix; HPA tissue IHC: profile; standard IHC practice). | Compare tissue structure and a no-primary control; adjust blocking and washing only if background impairs interpretation (standard IHC practice). |
| A salivary glandular population looks much weaker than esophageal epithelium. | The reported levels differ: Low in salivary glandular cells and High in esophageal squamous cells (HPA tissue IHC: cell-specific levels). | Score each annotated population against its own reference level; retain a known-positive control to judge assay performance (HPA tissue IHC: levels; standard IHC practice). |
| Strong staining appears in an HPA Not detected cell population but the positive control works. | HPA annotations are cell-specific and its reliability note flags staining in unannotated cells or structures; the signal needs review (HPA tissue IHC: negative list and reliability description). | Inspect morphology and HPA reference images, then compare an independent IHC-validated antibody if the assignment matters (HPA tissue IHC: reliability description; HPA antibodies: three Enhanced IHC entries). |
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 expect staining in respiratory epithelial cells (HPA: High in bronchus respiratory epithelial cells). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected); on the bronchus slide, morphologically identified cells without specific signal should show only background staining, without assuming a particular cell type is negative (standard IHC practice).
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 a no-primary, secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s clonality when known, and a validated ANXA2 knockout or immunizing-peptide competition control (selected-SKU IHC caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and check the bronchus control slide for background in inflammatory cells before interpreting DAB staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the PA1348 paraffin-section caption does not state the fixative (selected-SKU IHC caption). Heat retrieval in EDTA at pH 8.0 was used in that caption, but whether ANXA2 staining depends on retrieval or is easier in frozen sections is unreported (selected-SKU IHC caption). IF/ICC images exist for A-431, U-251MG and U2OS, with ANXA2 mainly at the plasma membrane and additionally in the cytosol; these data do not establish that IF is easier than IHC, while bronchial inflammatory cells warrant a check for endogenous peroxidase background (HPA subcellular; 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
Use compartment, cell type and slide controls together when troubleshooting ANXA2 staining in paraffin sections (HPA tissue IHC; HPA subcellular).
What should I adjust when ANXA2 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for this antibody (datasheet PA1348). Before changing retrieval, compare sections processed together using the reported 2 μg/ml primary concentration and overnight incubation at 4°C (PA1348 tissue-IHC caption). Check whether expected membranous staining appears in an appropriate positive cell population, such as liver cholangiocytes, while reviewing tissue preservation and background (HPA: High in liver cholangiocytes; HPA: ubiquitous membranous and extracellular expression; standard IHC practice). If signal remains weak, vary heating duration on matched sections while keeping the EDTA buffer and pH 8.0 fixed, then judge signal against tissue damage and background (standard IHC practice; datasheet PA1348).
Could fixation explain weak or uneven ANXA2 staining in paraffin sections?
ANXA2-specific sensitivity to fixation is unknown from the supplied evidence, and the PA1348 tissue-IHC caption does not state a fixative (PA1348 tissue-IHC caption). Record the actual fixation conditions and compare sections with similar processing histories before attributing uneven staining to the antibody (standard IHC practice). Use matched sections and the reported EDTA retrieval at pH 8.0 to assess whether staining tracks processing differences (datasheet PA1348; standard IHC practice). Assess morphology alongside signal, because poorly preserved tissue can make compartment-level scoring unreliable even when chromogen is visible (standard IHC practice). HPA staining patterns and ANXA2 modification annotations do not establish a fixation response for this antibody (HPA tissue IHC; UniProt P07355 modified residues).
Which ANXA2 staining compartments should I expect in chromogenic IHC?
Expect predominantly membranous staining, with possible extracellular signal, when reading tissue sections (HPA: ubiquitous membranous and extracellular expression). A cytosolic component is also plausible because approved subcellular observations place ANXA2 mainly at the plasma membrane and additionally in the cytosol (HPA subcellular). UniProt also annotates extracellular matrix and basement membrane localisation, without a transmembrane segment, so a cell-border pattern alone does not identify which side contains the epitope (UniProt P07355 localisation and topology). Score membrane, cytoplasm and extracellular deposits separately, then compare their distribution with intact tissue architecture and the expected stained cell population (HPA tissue IHC; standard IHC practice).
Can this stain distinguish ANXA2 isoforms or modified epitopes?
Do not assign a positive IHC stain to a particular ANXA2 isoform without antibody epitope mapping and isoform-specific validation (UniProt P07355: 2 isoforms; standard IHC practice). The supplied record lists modification sites, including SRC-associated phosphotyrosine at residue 24 and PKC-associated phosphoserine at residue 26, but it does not establish whether PA1348 recognizes either modified form (UniProt P07355 modified residues; PA1348 tissue-IHC caption). If staining differs between samples, first compare retrieval, morphology and compartment distribution under matched conditions (datasheet PA1348; standard IHC practice). Describe the result as antibody-detected ANXA2 staining unless a mapped epitope and appropriate controls support a narrower claim (standard IHC practice).
How should I investigate an ANXA2 pattern with multiplex IF?
Use multiplex IF as a follow-up to chromogenic IHC, pairing ANXA2 with a validated marker for the cell population being assessed, such as a cholangiocyte marker in liver (HPA: High in liver cholangiocytes; standard IF practice). Choose fluorophores and imaging channels after checking unstained tissue for autofluorescence, and include single-stain controls to evaluate channel bleed-through (standard IF practice). Plan permeabilisation around the antibody’s mapped epitope: access to a cytosolic epitope may require it, whereas staining of an accessible cell-surface epitope may not (standard IF practice). ANXA2 lacks an annotated transmembrane segment, and its observed plasma-membrane localisation does not establish the antibody’s epitope orientation (UniProt P07355 topology; HPA subcellular).
What should I check when ANXA2 DAB staining appears widespread?
First compare the stained section with a no-primary control to identify signal from the detection reagents or endogenous peroxidase (standard IHC practice). Check peroxidase blocking and inspect whether DAB deposits follow intact cells and structures, because the reported PA1348 workflow uses a peroxidase-linked secondary and DAB (PA1348 tissue-IHC caption; standard IHC practice). Widespread staining is not automatically nonspecific: HPA describes ubiquitous membranous and extracellular ANXA2 expression (HPA tissue IHC). If background obscures those compartments, compare matched sections while adjusting primary concentration from the reported 2 μg/ml and keeping retrieval, detection and exposure consistent (PA1348 tissue-IHC caption; standard IHC practice).
How can I quantify ANXA2 IHC without masking compartment differences? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, then record membranous and cytoplasmic staining separately and note extracellular deposits (HPA tissue IHC; HPA subcellular; standard IHC practice). For cellular staining, report percent positive cells and an intensity-weighted H-score using the same thresholds across samples (standard IHC practice). For a defined compartment or region, report stained area or positive-cell density per mm², with the denominator based on viable tissue area or the relevant cell population (standard IHC practice). Keep section thickness, imaging settings and scoring rules consistent, and exclude necrotic or damaged regions from the denominator according to a stated rule (standard IHC practice).
How do I distinguish credible ANXA2 staining from an artefact?
Judge a positive result by its location and cell identity: membranous or extracellular signal is consistent with tissue profiling, while cytosolic signal is also documented in subcellular observations (HPA tissue IHC; HPA subcellular). Check the named cell population where available, such as liver cholangiocytes, rather than treating every stained structure as equivalent (HPA: High in liver cholangiocytes; standard IHC practice). Reconsider staining confined to section edges, damaged areas or necrosis, and compare it with an intact region and a no-primary control (standard IHC practice). Diffuse DAB in a no-primary control points to a detection or endogenous-enzyme problem, while a reproducible, anatomically coherent pattern supports interpretation of the primary stain (standard IHC practice).
Boster reagents
Best ANXA2 / Annexin A2 IHC Antibodies
The anti-ANXA2 IHC image shows a human paraffin-embedded liver cancer section (PA1348 image caption). Catalog reactivity includes Human, Mouse and Rat (PA1348 catalog).
Real IHC data
Anti-Annexin A2/ANXA2 Antibody ®
Cat # PA1348
PA1348 is the only rendered card; its image shows chromogenic IHC in a paraffin-embedded human liver cancer section (PA1348 image caption). PA1348 lists Human, Mouse and Rat reactivity, while its supplied image documents human tissue only (PA1348 catalog; PA1348 image caption).
Which to pick: Choose PA1348 for tissue IHC: its own caption documents EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and HRP/DAB detection in a paraffin-embedded human liver cancer section; the fixative is unreported (PA1348 image caption). For IF/ICC, M00868 lists those applications and is a rabbit monoclonal, although no IF image is supplied (M00868 catalog). For cross-species work, both list Human, Mouse and Rat reactivity, but the supplied IHC image supports human tissue only (PA1348 catalog; M00868 catalog; 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 paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Secreted variants may separate RNA and protein locations (HPA tissue IHC); Isoform / epitope: Two isoforms; variant-specific epitope coverage unknown (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, PMC6694070. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANXA2/immunohistochemistry-ihc-guide.