ANXA13 / Annexin A13 · IHC design guide

Design Immunohistochemistry for ANXA13

Plan chromogenic ANXA13 IHC on paraffin sections with the catalog antibody at 1:50–1:200 (datasheet A11738). Use strongly stained small-intestine enterocytes as a positive reference and unstained adipocytes as a negative reference (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for ANXA13 (IHC for ANXA13): expected localisation Cytoplasmic staining in gastrointestinal, biliary, renal and tubal cells (HPA tissue IHC), antibody A11738, validated IHC image, and IHC protocol steps
Printable ANXA13 IHC protocol sheet — expected localisation Cytoplasmic staining in gastrointestinal, biliary, renal and tubal cells (HPA tissue IHC), antibody A11738, controls and protocol steps. Open the full ANXA13 IHC guide →

ANXA13 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 Cytoplasmic staining in gastrointestinal, biliary, renal and tubal cells (HPA tissue IHC)
Staining pattern Cytoplasmic signal in GI, bile duct, gallbladder, renal and tubal cells (HPA tissue IHC)
Antigen retrieval Citrate pH 6.0 HIER, heat-mediated (datasheet A11738)
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 across sections (standard IHC practice; not target-specific)
Caveat Signal varies by cell type within an organ (HPA tissue IHC)
Regulation RNA is enhanced in gallbladder and intestine (HPA tissue RNA)
Isoform / epitope 2 isoforms (A, B); epitope coverage is unknown (UniProt)
Section 1

Recommended ANXA13 IHC & IF Protocols

The catalog antibody protocol (datasheet A11738) is paired with published ANXA13 IHC methods for kidney sections (PMC12915081) and colorectal tumor and adjacent normal tissues (PMC5400614).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A11738)
FixationImage fixative and duration unreported (datasheet A11738); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: Citrate pH 6.0 (datasheet A11738); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibodyRabbit anti-ANXA13, 1:50-1:200 (datasheet A11738)
Primary incubationOvernight at 4 °C (standard)
DetectionHRP-polymer secondary, DAB chromogen 5–10 min (standard)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultANXA13-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in gastrointestinal tract, bile ducts, gallbladder, renal tubules and fallopian tube. No signal in the no-primary control.
💡Decision noteStart with heat-mediated citrate retrieval at pH 6.0 (datasheet A11738); the published methods specify microwave retrieval (PMC12915081) or heat retrieval without conditions (PMC5400614).
Section 2

What Is the Expected ANXA13 Staining Pattern?

ANXA13 is associated with the apical cell membrane and cytoplasmic vesicles; it has no transmembrane segment, and membrane association includes myristoylation and reversible calcium dependent lipid binding (UniProt P27216 topology and subcellular location). In paraffin IHC, expect epithelial staining in gastrointestinal tissue, gallbladder, bile ducts, renal tubules, and fallopian tube (HPA: tissue IHC profile). HPA rates its tissue pattern Enhanced, reflecting high consistency between staining and RNA expression (HPA: reliability).

What am I looking at on my slide?
Apical staining in small intestinal enterocytes, particularly the microvilli, with epithelial cytoplasmic staining.This fits the high enterocyte microvillar signal and broader gastrointestinal cytoplasmic profile observed by HPA (HPA: small intestine High; tissue IHC profile). UniProt also places ANXA13 at the apical membrane and in cytoplasmic vesicles (UniProt P27216 subcellular location). Judge the labeled cells and compartment together; uniform staining of every cell is not the expected tissue pattern (HPA: tissue IHC profile).
Staining is predominantly nuclear, while expected epithelial cytoplasmic or apical staining is absent.Treat this as discordant with the tissue IHC profile and investigate staining specificity or background (HPA: tissue IHC profile; standard IHC interpretation). A nuclear signal cannot automatically be dismissed across applications: HPA reports an additional, approved nucleoplasmic location in ICC-IF, alongside its supported plasma membrane location (HPA: subcellular ICC-IF).
Strong signal appears chiefly in cell types outside the reported positive populations.Compare cell identity with the documented pattern before scoring the section positive (HPA: tissue IHC positive-cell entries). For example, HPA reports high signal in duodenal endocrine cells and medium signal in colonic goblet cells, which are different populations (HPA: duodenum High; colon Medium). Unexpected staining can reflect antibody cross-reactivity or endogenous detection activity; it does not establish ANXA13 expression without supporting controls (standard IHC interpretation).
Diffuse color covers tissue and spaces between cells, obscuring epithelial boundaries.This distribution is difficult to reconcile with the reported cell-associated cytoplasmic and apical pattern (HPA: tissue IHC profile; UniProt P27216 subcellular location). Consider nonspecific antibody binding, incomplete blocking, or excess chromogen development, and inspect the negative reagent control (standard IHC practice). Background intensity alone should not be scored as target-positive staining (standard IHC interpretation).
No signal appears in a section containing expected positive epithelial cells.An absence in small intestinal enterocytes or appendix or duodenal endocrine cells conflicts with HPA's high staining in those specified cells (HPA: small intestine, appendix, and duodenum High). First verify that the relevant cells are present and preserved; then review the IHC-validated antibody, detection reagents, and staining conditions with a positive control (standard IHC practice). A negative slide by itself cannot establish absence of ANXA13 (standard IHC interpretation).
💡Expected ANXA13 appearanceCall a result positive when the expected epithelial cells show clear cytoplasmic or apical staining, including high microvillar signal in small intestinal enterocytes; widespread signal in unrelated cells or structureless background is suspect (HPA: tissue IHC profile; small intestine High; standard IHC interpretation).
How each factor affects the staining
Cell type and siteExpected intensity varies by population: small intestinal enterocyte microvilli and fallopian tube cilia axonemes are High, whereas colonic goblet cells, gallbladder glandular cells, and kidney proximal tubule microvilli are Medium (HPA: tissue IHC positive-cell entries). Score the named cells, not the whole organ as one uniform compartment (standard IHC interpretation).
Membrane association and topologyANXA13 lacks a transmembrane segment, yet myristoylation anchors it to membranes and calcium dependent binding can be reversible (UniProt P27216 topology and subcellular location). This supports interpreting apical or membrane-associated staining alongside cytoplasmic vesicle signal; it does not prescribe an antigen-retrieval condition for paraffin sections (UniProt P27216 subcellular location; standard IHC practice).
Antibody validationHPA lists Enhanced IHC validation for HPA018535, HPA019569, HPA019650, and CAB025135; its tissue reliability is also Enhanced (HPA: antibodies; tissue IHC reliability). These summaries support the reported pattern but do not make an unexpected compartment or cell population positive without controls (standard IHC interpretation).
IF/ICC Q: What localization should an IF result show?A: HPA reports mainly plasma membrane localization, supported in ICC-IF, with additional approved nucleoplasmic localization; image-bearing cell lines are A-549 and U-251MG (HPA: subcellular ICC-IF). That ICC-IF observation is separate from the paraffin tissue IHC pattern and supplies no IHC protocol condition (HPA: subcellular ICC-IF; tissue IHC profile).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Expected-positive epithelium is unstained.The relevant cells may be absent from the section, or the IHC detection run may have failed; HPA reports high staining only for specified positive cells, such as small intestinal enterocyte microvilli (HPA: small intestine High; standard IHC practice).Confirm the cells on the counterstained section, then compare a known-positive section processed in the same run; review the catalog antibody's IHC-P instructions and reagent performance (standard IHC practice).
Color is diffuse across cells and extracellular areas.Nonspecific binding, inadequate blocking, or excessive development can obscure the cell-associated pattern (standard IHC practice; HPA: tissue IHC profile).Inspect a negative reagent control; optimize blocking, antibody dilution, washing, and chromogen development according to the IHC workflow (standard IHC practice).
Staining is mainly nuclear in tissue IHC.The tissue IHC pattern is described as cytoplasmic, although HPA separately approves an additional nucleoplasmic ICC-IF location (HPA: tissue IHC profile; subcellular ICC-IF).Check whether epithelial cytoplasmic or apical staining is also present, inspect controls, and avoid assigning isolated nuclear color to ANXA13 without corroboration (standard IHC interpretation).
Strong signal appears in an unexpected cell population.Cross-reactivity or endogenous detection activity is possible when staining does not follow the documented cell-specific pattern (HPA: tissue IHC positive-cell entries; standard IHC interpretation).Identify the stained cells with the counterstain, inspect reagent controls, and compare with the named HPA positive and not-detected populations before scoring (HPA: tissue IHC positive and negative entries; standard IHC practice).
A low-signal tissue is being used to judge assay failure.HPA records Low staining in nasopharyngeal and bronchial ciliated rootlets and stomach glandular cells, so faint signal there has limited value as a positive-run check (HPA: tissue IHC low entries).Use a section containing a documented High population, such as small intestinal enterocyte microvilli, as the positive comparison; score the specified cells (HPA: small intestine High; standard IHC practice).
A not-detected tissue shows convincing-looking color.HPA reports Not detected in specified cells, including adipocytes in adipose tissue and hematopoietic cells in bone marrow; apparent color warrants specificity review (HPA: tissue IHC negative entries).Verify cell identity and inspect negative reagent controls, then compare staining with a documented positive cell population in the same run (HPA: tissue IHC positive and negative entries; standard IHC practice).

Sample controls for ANXA13 IHC & IF

🧪Run small intestine first: enterocyte microvilli should stain strongly (HPA: High in small-intestine enterocytes). Use adipose tissue as the negative: adipocytes are not detected (HPA: Not detected in adipocytes). On the positive slide, assess surrounding non-enterocyte cells for background rather than assuming they are ANXA13-negative; the supplied HPA row scores enterocytes only (HPA: small-intestine enterocytes).
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 ANXA13 in A-549, U-251MG, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; match an isotype control to the primary antibody’s host and, for a monoclonal antibody, its isotype; and use ANXA13-knockout tissue as a biological negative if available. For chromogenic IHC, quench endogenous peroxidase and check background in the small-intestine section (HPA: High in small-intestine enterocytes); block endogenous biotin if using biotin-based detection.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected A11738 paraffin-section caption is unreported (selected-SKU tissue-IHC caption: fixative not stated). The caption uses high-pressure retrieval in 10 mM citrate at pH 6.0 with a 1:50 dilution in colon carcinoma; it provides a starting condition but does not establish retrieval dependency for small intestine (selected-SKU tissue-IHC caption). Frozen-section ease is unreported; ICC-IF images exist for A-549 and U-251MG (HPA: ICC-IF image cell lines), while intestinal luminal contents can complicate interpretation of apical staining (HPA: High in enterocyte microvilli).

HPA tissue IHC evidence for ANXA13

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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

TissueCell typeLevelEvidenceSource
Appendix Endocrine cells High Protein (IHC) HPA →
Duodenum Endocrine cells High Protein (IHC) HPA →
Fallopian tube Ciliated cells (cilia axoneme) High Protein (IHC) HPA →
Small intestine Enterocytes - Microvilli High Protein (IHC) HPA →
Testis Sertoli cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Not detected Protein (IHC) HPA →
Adrenal gland Glandular cells Not detected Protein (IHC) HPA →
Bone marrow Hematopoietic cells Not detected Protein (IHC) HPA →
Breast Adipocytes Not detected Protein (IHC) HPA →
Caudate Glial cells Not detected Protein (IHC) HPA →
Section 3

Advanced ANXA13 IHC Tips

Troubleshoot ANXA13 staining in paraffin sections by checking retrieval, cell identity and subcellular pattern before comparing staining intensity.

What should I change when ANXA13 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in 10 mM citrate buffer at pH 6.0 (datasheet A11738). The A11738 paraffin-section image used high-pressure retrieval and a 1:50 antibody dilution, giving a directly documented starting condition for this antibody (caption A11738). If staining is weak, check that sections were fully deparaffinised and that heating and cooling were consistent across the batch before changing retrieval chemistry (standard IHC practice). Compare a retrieval adjustment on adjacent sections alongside a known positive intestinal epithelial control; preserve the same detection and counterstain settings so any change in signal remains interpretable (UniProt P27216 tissue specificity; standard IHC practice).
Could fixation explain a weak or patchy ANXA13 result?
Target-specific fixation sensitivity for ANXA13 is unknown from the supplied evidence, and the A11738 paraffin-section caption does not state its fixative (caption A11738). Record the actual fixative, fixation duration and tissue thickness for each specimen before comparing staining across cases (standard IHC practice). Process a positive control with the study material where possible, then compare adjacent sections using the same citrate retrieval and detection settings (UniProt P27216 tissue specificity; datasheet A11738; standard IHC practice). If staining differs by batch, assess fixation history and section quality alongside antibody and detection controls; an IHC pattern alone cannot establish a fixation effect specific to ANXA13 (standard IHC practice).
Should ANXA13 look membranous or cytoplasmic in a tissue section?
Assess epithelial staining at the apical membrane and in cytoplasmic compartments: ANXA13 is annotated at the apical cell membrane, cell membrane and cytoplasmic vesicles (UniProt P27216 subcellular). Its membrane association includes myristoylation and reversible calcium-dependent lipid binding, while its sequence has no transmembrane segment (UniProt P27216 subcellular and topology). Tissue IHC also reports cytoplasmic expression, with high staining in small-intestinal enterocyte microvilli and fallopian-tube ciliated-cell axonemes (HPA tissue IHC). Check these patterns against tissue architecture and counterstained cell borders; a diffuse stain without an identifiable epithelial compartment needs review of background and control sections (HPA tissue IHC; standard IHC practice).
Can an unknown epitope make one ANXA13 staining pattern misleading?
ANXA13 has 2 recorded isoforms, A and B, so establish which sequence the antibody immunogen covers before assigning staining to either isoform (UniProt P27216 isoforms; standard IHC practice). The supplied A11738 caption documents tissue staining and retrieval, but provides no epitope or isoform specificity (caption A11738). An extracellular-epitope assumption would be unsupported because ANXA13 has no transmembrane segment and is associated with membranes through myristoylation and calcium-dependent binding (UniProt P27216 topology and subcellular). If isoform attribution matters, compare the immunogen sequence with both isoforms and use an independent isoform-resolving assay; ordinary chromogenic IHC intensity alone cannot make that assignment (standard IHC practice).
How can IF help resolve an ambiguous ANXA13 IHC pattern?
Use IF/ICC as a separate validation experiment, multiplexing ANXA13 with a marker that identifies the expected epithelial cell population in the specimen (UniProt P27216 tissue specificity; standard IF practice). Select fluorophores after inspecting unstained tissue and place the weaker channel away from the strongest tissue autofluorescence; include single-stain controls when channels may overlap (standard IF practice). For an epitope on the cytoplasmic face of a membrane or within vesicles, optimise gentle permeabilisation against membrane preservation, since ANXA13 has no transmembrane segment and its annotated locations include membranes and cytoplasmic vesicles (UniProt P27216 topology and subcellular; standard IF practice). Compare cell identity and apical distribution with the chromogenic section, without assuming the two detection methods have identical sensitivity (standard IHC/IF practice).
How do I reduce background without losing apical ANXA13 staining?
First distinguish diffuse reagent background from structured staining at epithelial apical surfaces, because ANXA13 is annotated at the apical membrane and HPA reports enterocyte microvillar staining (UniProt P27216 subcellular; HPA tissue IHC). Include a no-primary control, inspect endogenous pigment, and assess nonspecific secondary or detection-reagent staining before lowering antibody concentration (standard IHC practice). The documented A11738 image used 1:50; titrate around that condition on matched sections while holding retrieval and development constant (caption A11738; standard IHC practice). For peroxidase-based chromogenic detection, apply an endogenous-peroxidase block and control DAB development time, then judge whether the epithelial pattern remains above the control background (standard IHC practice).
What should I score when ANXA13 signal varies within epithelium? ⚠ ANSWER MARKED FOR VERIFICATION
Define the epithelial region and cell type before scoring, because ANXA13 is detected in colon and jejunum epithelium and HPA identifies cell-specific staining across tissues (UniProt P27216 tissue specificity; HPA tissue IHC). Record the percentage of positive target cells and intensity, then calculate an H-score if intensity categories are reproducible; report apical and cytoplasmic staining separately when distinguishable (standard IHC practice; UniProt P27216 subcellular). Normalise to the number of eligible epithelial cells or epithelial area, and use density per mm² only when the sampled area is defined consistently (standard IHC practice). Keep retrieval, exposure to chromogen and scoring thresholds consistent across cases, with blinded review for borderline staining (standard IHC practice).
Which findings support a true ANXA13-positive IHC result?
A credible result follows epithelial anatomy and the expected apical or cytoplasmic distribution, with a matching positive control; intestinal epithelial expression and apical membrane association are documented for ANXA13 (UniProt P27216 tissue specificity and subcellular). HPA reports high enterocyte microvillar staining in small intestine and medium goblet-cell staining in colon, so judge each cell population against its own context (HPA tissue IHC). Question staining confined to cut edges or necrotic areas, or staining that appears equally in the no-primary control; these patterns indicate possible section or detection artefact (standard IHC practice). For peroxidase detection, investigate residual endogenous enzyme signal before calling unexpected cells positive (standard IHC practice).
Boster reagents

Best ANXA13 / Annexin A13 IHC Antibodies

A11738 has IHC images from paraffin-embedded human colon carcinoma and mouse jejunum (catalog IHC captions). Human, mouse and rat reactivity is listed; no IF image is supplied (catalog reactivity; catalog images).

Real IHC data Immunohistochemistry of paraffin-embedded human colon carcinoma using ANXA13 antibody at dilution of 1:50 .Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.
Anti-Annexin A13 ANXA13 Antibody
Cat # A11738

A11738 is listed for IHC and has paraffin-section images from human colon carcinoma and mouse jejunum, each stained at 1:50 after high-pressure retrieval in 10 mM citrate, pH 6.0 (catalog applications; catalog IHC captions). Its listed reactivity is human, mouse and rat; the payload supplies no IF image (catalog reactivity; catalog images).

Which to pick: Choose A11738 for paraffin-section IHC: its own captions document staining in human colon carcinoma and mouse jejunum, but do not report the fixative (catalog IHC captions). For cross-species work, A11738 lists human, mouse and rat reactivity; the supplied images document human and mouse tissue only (catalog reactivity; catalog IHC captions). For IF/ICC, A11738 has no listed IF application or IF image, although its catalog text mentions immunofluorescence microscopy testing; optimize and verify it for the intended sample (catalog applications; catalog text; catalog images).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry P27216 (ANX13_HUMAN, Annexin A13).
  2. Human Protein Atlas. ANXA13 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. ANXA13 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the nucleoplasm..
  4. Human Protein Atlas. ANXA13 antibody validation summary (4 antibodies).
  5. Annexin A13 Protects Against Acute Kidney Injury by Inactivating TGF-β/Smad3 Signaling. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC12915081.
  6. ANXA13 expression patterns in hepatocellular carcinoma and impact on tumor behavior. World journal of gastrointestinal surgery 2025 — PMC12576647.
  7. Annexin Expression in Cholangiocarcinoma, and Metastatic Pancreatic Ductal Adenocarcinoma "Is it be Helpful for Differential Diagnosis of These Tumors in the Liver?". Iranian journal of pathology 2021 — PMC8463754.
  8. Annexin A13 promotes tumor cell invasion in vitro and is associated with metastasis in human colorectal cancer. Oncotarget 2017 — PMC5400614.
  9. PubMed PMID:1530946 — UniProt-cited evidence.
  10. PubMed PMID:11961095 — UniProt-cited evidence.
  11. PubMed PMID:16421571 — UniProt-cited evidence.