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ANXA13 / Annexin A13 · IHC design guide
Design Immunohistochemistry for ANXA13
Plan ANXA13 staining in paraffin sections using cytoplasmic expression in gastrointestinal, biliary, renal and fallopian tissues as a reference (HPA tissue IHC). The catalog antibody has an IHC dilution range of 1:50–1:200 (datasheet A11738); use cell-specific controls when interpreting the stain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
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 tissue stain (HPA tissue IHC); apical membrane association (UniProt) | |
| Staining pattern | Cytoplasmic staining in GI, biliary, renal and fallopian tissues (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11738) | |
| Caveat | GI endocrine cells stain high; colon goblet cells medium (HPA tissue IHC) | |
| Regulation | Tissue enhanced in gallbladder and intestine (HPA tissue RNA) | |
| Isoform / epitope | Two isoforms, A and B (UniProt); antibody epitope unspecified (datasheet A11738) |
Section 1
Recommended ANXA13 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A11738) with published ANXA13 IHC protocols for kidney sections (PMC12915081) and colorectal tissue (PMC5400614).
| Sample | Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A11738) |
| Fixation | Image fixative and duration unreported (datasheet A11738); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A11738); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-ANXA13, 1:50-1:200 (datasheet A11738) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ANXA13-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 citrate pH 6.0 heat retrieval (datasheet A11738). The published protocols report microwave or heat retrieval without specifying a buffer (PMC12915081; PMC5400614).
Section 2
What Is the Expected ANXA13 Staining Pattern?
ANXA13 is associated with the apical cell membrane, cell membrane and cytoplasmic vesicles, and has no transmembrane segment (UniProt P27216 topology). In paraffin-section IHC, expect staining in selected epithelial cells, especially intestinal enterocytes; HPA describes a broader cytoplasmic gastrointestinal and ductal profile with Enhanced tissue-IHC reliability, reflecting consistency between antibody staining and RNA expression (HPA tissue IHC: Enhanced; HPA tissue IHC: enterocytes, High).
| Apical or cytoplasmic staining in small-intestinal enterocytes, especially along microvilli. | This fits a known strong site: HPA scores enterocyte microvilli High (HPA tissue IHC: small intestine, High). Apical membrane association and vesicles also fit the protein annotation (UniProt P27216: subcellular location). Assess the epithelial pattern and tissue structure together; intensity alone cannot establish antibody specificity (general IHC practice). |
| Predominantly nuclear staining in intestinal epithelium, with little epithelial apical or cytoplasmic signal. | Treat this as discordant with the reported tissue-IHC pattern (HPA tissue IHC: cytoplasmic profile). Check morphology and controls before calling it ANXA13 (general IHC practice). HPA reports additional nucleoplasmic localisation in ICC-IF, so nuclear fluorescence in that separate application is not, by itself, evidence of an IHC artefact (HPA subcellular: nucleoplasm, approved). |
| Strong signal in adipocytes or hematopoietic cells, especially when expected epithelial staining is absent. | These cell populations are reported as Not detected in the sampled adipose tissue and bone marrow, respectively (HPA tissue IHC: adipocytes, Not detected; HPA tissue IHC: hematopoietic cells, Not detected). Review cell identification and negative controls; off-target binding or endogenous detection activity are possible explanations, not diagnoses from staining alone (general IHC practice). |
| A uniform chromogenic haze covers epithelium, stroma and blank areas, obscuring cell boundaries. | A field-wide haze is difficult to reconcile with HPA's cell-selective tissue pattern (HPA tissue IHC: cell-specific levels). Interpret the apparent ANXA13 signal only after checking reagent background, blocking, washes and the no-primary control (general IHC practice). Do not score the haze as cytoplasmic positivity simply because HPA describes cytoplasmic expression (HPA tissue IHC: profile). |
| No signal appears in well-preserved small-intestinal enterocytes. | This conflicts with HPA's High enterocyte-microvillus observation (HPA tissue IHC: small intestine, High). First verify that the expected cells are present and that the detection run worked (general IHC practice). A negative result can reflect assay performance or specimen variation; the supplied sources do not establish a target-specific fixation effect (HPA tissue IHC; UniProt P27216). |
💡Expected ANXA13 appearanceCall a positive result when epithelial staining is interpretable at enterocyte microvilli or the apical/cytoplasmic region, with potentially High small-intestinal enterocyte signal (HPA tissue IHC: small intestine, High; UniProt P27216: apical membrane); diffuse haze or strong staining in HPA Not detected cell populations is suspect and needs controls (HPA tissue IHC: negative cells; general IHC practice).
| Cell and tissue selection | HPA reports High signal in small-intestinal enterocyte microvilli, appendix and duodenum endocrine cells, fallopian-tube ciliated-cell axonemes and testis Sertoli cells; colon goblet cells, gallbladder glandular cells and kidney proximal-tubule microvilli are Medium (HPA tissue IHC). A negative cell type within a tissue does not make the entire tissue a negative control (HPA tissue IHC: cell-specific levels). |
| Membrane association | ANXA13 lacks a transmembrane segment, is anchored by myristoylation and also binds lipid membranes reversibly in a calcium-dependent manner (UniProt P27216: topology and subcellular annotation). Membrane-associated staining can therefore coexist with cytoplasmic or vesicular signal; these annotations do not predict a fixation or retrieval response (UniProt P27216; HPA tissue IHC: cytoplasmic profile). |
| Isoforms and antibody evidence | UniProt lists isoforms A and B (UniProt P27216: isoforms). The supplied record gives no epitope mapping or isoform-specific IHC pattern, so staining cannot be assigned to either isoform from location alone. Four listed antibodies have Enhanced IHC status (HPA antibodies: HPA018535, HPA019569, HPA019650, CAB025135); that supports the reported pattern without proving every signal in a new specimen is specific. |
| IF/ICC Q&A | What localisation should IF/ICC show? HPA reports plasma membrane as the main, supported location and nucleoplasm as an additional, approved location (HPA subcellular: ICC-IF). Its images include A-549 and U-251MG; their listing does not establish an IHC-positive control or an IF staining intensity for either line (HPA subcellular: image cell lines). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected enterocyte signal is absent. | The positive cell population may be missing from the section, or the IHC run may have failed (general IHC practice); HPA reports High small-intestinal enterocyte staining (HPA tissue IHC). | Confirm enterocyte morphology, inspect a known-positive section and check the detection controls before interpreting the sample as negative (general IHC practice). |
| Signal is mostly nuclear in paraffin IHC. | The distribution differs from HPA's cytoplasmic tissue-IHC profile, although ICC-IF has an additional nucleoplasmic location (HPA tissue IHC: profile; HPA subcellular: nucleoplasm). | Review tissue morphology and control slides, then repeat or compare with another IHC-validated antibody if the result remains consequential (general IHC practice; HPA antibodies: IHC Enhanced). |
| Diffuse chromogenic background obscures cell boundaries. | Nonspecific reagent deposition or endogenous detection activity can obscure a cell-specific pattern (general IHC practice; HPA tissue IHC: cell-specific levels). | Inspect no-primary and detection controls; review blocking, washes and detection conditions before scoring compartments (general IHC practice). |
| An HPA Not detected cell population stains strongly. | Cell misidentification, cross-reactivity or endogenous detection activity are possibilities (general IHC practice; HPA tissue IHC: Not detected cell populations). | Recheck the named cell type and compare a negative-control section; avoid labeling the entire tissue positive or negative from one cell population (general IHC practice; HPA tissue IHC: cell-specific levels). |
| Only weak signal appears in a proposed control tissue. | HPA reports Low staining in stomach glandular cells and ciliated cells of bronchus or nasopharynx, so weak staining there may match the reference pattern (HPA tissue IHC: Low). | For a clearer positive comparator, use a section containing small-intestinal enterocyte microvilli, reported High, and score the specified cells (HPA tissue IHC: small intestine, High). |
| Apical and cytoplasmic signals differ between cases. | Both compartments are consistent with reported localisation, but the supplied sources do not establish a target-specific fixation effect or explain case-to-case differences (UniProt P27216: subcellular location; HPA tissue IHC: profile). | Score apical and cytoplasmic signal separately against matched controls and tissue morphology; report unexplained differences without assigning them to fixation (general IHC practice). |
Experiment evidence
IHC & IF images showing ANXA13 staining
A11738 · IHCImmunohistochemistry of paraffin-embedded human colon carcinoma using ANXA13 antibody at dilution of 1:50 .Perform high pressure antigen ret
A11738 · IHCImmunohistochemistry of paraffin-embedded mouse jejunum using ANXA13 antibody at dilution of 1:50 .Perform high pressure antigen retrieval w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104537-ANXA13/tissue/cerebral+cortex#imid_18789300
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000104537-ANXA13/tissue/duodenum#imid_18789233
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104537-ANXA13/tissue/colon#imid_18789226
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104537-ANXA13/tissue/liver#imid_18789227
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA019650; green = ANXA13, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA019650; green = ANXA13, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ANXA13 IHC & IF
🧪Run small intestine first and require apical staining in enterocyte microvilli (HPA: High in small intestine enterocytes). Use adipose tissue adipocytes as the biological negative (HPA: Not detected in adipocytes); on the positive slide, assess non-enterocyte cells lacking apical signal as internal background, without treating them as validated ANXA13-negative cells (HPA: High assigned to enterocyte microvilli).
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, a host-species- and clonality-matched isotype control, and ANXA13-knockout tissue or cells as a biological specificity control (standard IHC practice). Block endogenous peroxidase for chromogenic detection and assess intestinal autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A11738 paraffin-section caption does not report a fixative (selected-SKU caption: fixative unreported). Start retrieval assessment with high-pressure 10 mM citrate, pH 6.0, the condition reported for colon carcinoma IHC at 1:50 (selected-SKU tissue-IHC caption); its necessity for small intestine is unreported. The supplied evidence does not establish whether frozen sections or IF are easier; check apical signal against intestinal luminal staining and background (HPA: High in enterocyte microvilli; standard IHC practice).
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| 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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| 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 compartment before comparing signal intensity (UniProt P27216; HPA tissue IHC).
What should I change if ANXA13 staining is weak after antigen retrieval?
Use heat-mediated retrieval in 10 mM citrate buffer, pH 6.0, as the starting condition for this antibody (datasheet A11738; A11738 tissue-IHC caption). The reported paraffin-section image used high-pressure retrieval followed by antibody at 1:50; its fixative was not stated (A11738 tissue-IHC caption). Confirm that sections received consistent heating and cooling, then compare a known positive intestinal section across runs using the same detection conditions (standard IHC practice; UniProt P27216 tissue specificity). If signal remains weak, test a longer heating interval on adjacent sections while watching for tissue damage; document each condition rather than attributing a change to ANXA13 alone (standard IHC practice).
How can I assess whether fixation is causing weak ANXA13 IHC staining?
Target-specific ANXA13 sensitivity to any fixative or fixation duration is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (A11738 tissue-IHC caption). Record fixative, fixation time and processing history for each block, then compare sections with a shared retrieval and detection run (standard IHC practice). Include a section with an expected intestinal epithelial signal to check whether the run can reveal ANXA13 under those conditions (UniProt P27216 tissue specificity; standard IHC practice). If only one block is weak, investigate its processing history and tissue preservation before assigning the difference to ANXA13 biology (standard IHC practice).
Where should I expect ANXA13 staining within positive tissue cells?
Assess both epithelial cytoplasm and the apical membrane region: ANXA13 is reported at the apical cell membrane and in cytoplasmic vesicles, while tissue IHC shows a cytoplasmic profile (UniProt P27216 subcellular location; HPA tissue IHC profile). Small-intestinal enterocyte microvilli show high staining, and colon goblet cells show medium staining in the supplied tissue observations (HPA tissue IHC). Compare the luminal border with neighboring cytoplasm at the same magnification, keeping the epithelial cell type identifiable (standard IHC practice). Do not require a continuous membrane rim in every cell, because ANXA13 also binds lipid membranes reversibly in a calcium-dependent manner (UniProt P27216 subcellular location).
Could ANXA13 isoforms or epitope position explain different staining patterns?
ANXA13 has 2 reported isoforms, A and B, and no annotated transmembrane segment; the supplied evidence does not map this antibody's epitope to either isoform (UniProt P27216 isoforms and topology; A11738 tissue-IHC caption). Check the antibody's documented immunogen or epitope, if available, before calling a compartment difference isoform-specific (standard IHC practice). On adjacent sections, keep retrieval and antibody concentration constant while comparing the apical region and cytoplasm (standard IHC practice; UniProt P27216 subcellular location). A change in staining after retrieval optimization establishes an assay effect, not isoform identity; isoform assignment needs independent isoform-specific evidence (standard IHC practice).
How should I investigate ANXA13 localisation by IF alongside the IHC result?
For a separate IF assay, multiplex ANXA13 with a marker that identifies the expected epithelial cell type, then inspect whether signal follows the apical border or intracellular structures (UniProt P27216 subcellular location; standard IF practice). Choose fluorophores after checking unstained tissue autofluorescence in each channel, and use single-stain controls to assess bleed-through (standard IF practice). ANXA13 has no transmembrane segment, but this antibody's epitope side is unspecified; determine epitope accessibility before deciding whether detergent permeabilisation is needed (UniProt P27216 topology; A11738 tissue-IHC caption; standard IF practice). Treat the paraffin-section citrate retrieval and 1:50 dilution as IHC evidence only when planning IF conditions (A11738 tissue-IHC caption).
How do I separate ANXA13 signal from diffuse chromogenic background?
Run a no-primary control on the same tissue and inspect whether diffuse color persists after the chromogenic detection sequence (standard IHC practice). Review blocking, washes, antibody concentration and chromogen development together; the selected paraffin-section image used 1:50, which is a reported condition rather than a universal optimum (standard IHC practice; A11738 tissue-IHC caption). Include a peroxidase block when using peroxidase-based DAB detection, and compare with a known positive epithelial region (standard IHC practice; UniProt P27216 tissue specificity). Call signal convincing when it resolves to expected cells and compartments above the control background, rather than following section edges or damaged areas (HPA tissue IHC; standard IHC practice).
What should I score when comparing ANXA13 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define a tissue compartment and cell population before scoring, such as enterocytes in small intestine or goblet cells in colon (HPA tissue IHC). Record the percentage of positive target cells and intensity, then calculate an H-score from 0–300 if intensity grading is reproducible (standard IHC practice). Score apical enrichment separately from diffuse cytoplasmic staining when that distinction addresses the study question (UniProt P27216 subcellular location; HPA tissue IHC profile). Normalize to the number of evaluable target cells or their tissue area, keep retrieval and imaging conditions consistent, and exclude folds, necrosis and poorly preserved regions (standard IHC practice).
When is an apparent ANXA13-positive IHC result likely to be artefactual?
Give greatest weight to staining in the expected epithelial cells and apical or cytoplasmic regions: intestinal epithelium is supported by protein-level evidence, and tissue IHC reports gastrointestinal cytoplasmic expression (UniProt P27216 tissue specificity and subcellular location; HPA tissue IHC profile). Isolated color in an unexpected cell type or compartment needs comparison with a known positive section and a no-primary control (standard IHC practice). Edge-only staining, necrotic deposits and color retained in the no-primary control point toward processing or detection artefact (standard IHC practice). Check endogenous peroxidase activity when using peroxidase-based DAB, and do not infer ANXA13 absence from one weakly stained block without checking its preservation and retrieval (standard IHC practice).
Boster reagents
Best ANXA13 / Annexin A13 IHC Antibodies
The IHC-validated ANXA13 antibody has images from paraffin-embedded human colon carcinoma and mouse jejunum; catalog reactivity also includes rat (A11738 IHC image captions; A11738 reactivity list).
Real IHC data
Anti-Annexin A13 ANXA13 Antibody
Cat # A11738
A11738 has IHC images of paraffin-embedded human colon carcinoma and mouse jejunum, each stained at 1:50 after high-pressure retrieval in 10 mM citrate, pH 6.0 (A11738 IHC image captions). Its listed applications include IHC, and its listed reactivity covers human, mouse and rat (A11738 applications and reactivity lists).
Which to pick: Choose A11738 for paraffin-section tissue IHC using the captioned retrieval conditions as a starting point; the fixative is unreported (A11738 IHC image captions). For IF/ICC, A11738 has no listed IF application or IF image, so establish staining conditions and specificity for that use (A11738 applications list; A11738 IF image list). A11738 is the only catalog choice here for cross-species work, with human, mouse and rat reactivity listed but IHC images shown only for human and mouse samples (A11738 reactivity list; A11738 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P27216 (ANX13_HUMAN, Annexin A13).
- Human Protein Atlas. ANXA13 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ANXA13 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the nucleoplasm..
- Human Protein Atlas. ANXA13 antibody validation summary (4 antibodies).
- Annexin A13 Protects Against Acute Kidney Injury by Inactivating TGF-β/Smad3 Signaling. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC12915081.
- ANXA13 expression patterns in hepatocellular carcinoma and impact on tumor behavior. World journal of gastrointestinal surgery 2025 — PMC12576647.
- 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.
- Annexin A13 promotes tumor cell invasion in vitro and is associated with metastasis in human colorectal cancer. Oncotarget 2017 — PMC5400614.
- PubMed PMID:1530946 — UniProt-cited evidence.
- PubMed PMID:11961095 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ANXA13 (Annexin A13). Expected localisation Cytoplasmic tissue stain (HPA tissue IHC); apical membrane association (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A11738). Validated IHC antibody: A11738 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet A11738); 2. Fixation: Image fixative and duration unreported (datasheet A11738); 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: Citrate pH 6.0 (datasheet A11738); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-ANXA13, 1:50-1:200 (datasheet A11738); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11738); Caveat: GI endocrine cells stain high; colon goblet cells medium (HPA tissue IHC); Isoform / epitope: Two isoforms, A and B (UniProt); antibody epitope unspecified (datasheet A11738); Regulation: Tissue enhanced in gallbladder and intestine (HPA tissue RNA). Evidence: UniProt P27216, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A11738, PMC12915081, PMC5400614. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANXA13/immunohistochemistry-ihc-guide.