This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ANXA7 / Annexin A7 · IHC design guide
Design Immunohistochemistry for ANXA7
Plan ANXA7 paraffin IHC using the reported cytoplasmic and membranous tissue pattern (HPA tissue IHC). Start with the catalog antibody's 0.5–1 μg/mL IHC range (datasheet PA2076), and interpret staining alongside HPA's low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ANXA7 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Broad cytoplasmic and membranous staining; glandular cells positive (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PA2076) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+1 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 | Antibody staining and RNA levels show low consistency (HPA tissue IHC) | |
| Regulation | Isoform abundance differs by tissue (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended ANXA7 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet: PA2076). Two published ANXA7 IHC protocols provide additional paraffin-section conditions (PMC6188866; PMC11659777).
| Sample | Paraffin-embedded Human Mammary Cancer tissue; fixative not specified (datasheet PA2076) |
| Fixation | Image fixative and duration unreported (datasheet PA2076); 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 PA2076); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PA2076) |
| Primary antibody | Rabbit anti-ANXA7, 0.5-1μg/ml (datasheet PA2076) |
| Primary incubation | Overnight at 4 °C (datasheet PA2076) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PA2076) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ANXA7-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Ubiquitous cytoplasmic and membranous expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: PA2076). The published protocols used microwave retrieval; only one specifies sodium citrate buffer (PMC6188866; PMC11659777).
Section 2
What Is the Expected ANXA7 Staining Pattern?
ANXA7 should appear mainly in cytoplasm and along cell membranes, with medium staining reported in several glandular and respiratory epithelial cells and in hematopoietic and lymphoid tissue (HPA: ubiquitous cytoplasmic and membranous expression; listed medium signals). It has no transmembrane segment, so membrane staining need not form a continuous surface outline (UniProt P20073: no transmembrane segment). HPA rates its tissue IHC Approved while noting low consistency with RNA expression (HPA: reliability description).
| Cytoplasmic staining, with some membranous accentuation, in adrenal or breast glandular cells. | This fits the reported compartment and cell-level pattern; HPA lists medium staining in those cells (HPA: tissue IHC). Judge intensity against a matched control section, since slide preparation and detection affect its appearance (general IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic or membranous signal. | That distribution does not match HPA’s reported cytoplasmic and membranous pattern (HPA: tissue IHC). Treat it as unconfirmed; inspect the negative control and repeat with adjusted staining conditions before assigning ANXA7 to nuclei (general IHC practice). |
| Strong adipocyte or small-intestinal glandular-cell staining. | HPA reports ANXA7 as not detected in those respective cells (HPA: tissue IHC). Check morphology and background; unexpected signal may reflect cross-reactivity or endogenous detection activity, but this pattern alone does not establish its cause (general IHC practice). |
| Broad, hazy colour across cells, stroma and empty areas. | Signal without a recognizable cellular pattern is difficult to score as ANXA7. Compare a no-primary control and check wash, blocking and detection steps for background (general IHC practice); HPA’s expected pattern is cytoplasmic and membranous (HPA: tissue IHC). |
| No staining in a technically adequate section of adrenal gland or breast. | Their glandular cells are reported at medium intensity, so complete absence warrants a control and workflow check (HPA: tissue IHC). It does not by itself prove the specimen lacks ANXA7; confirm the tissue and cell type before interpreting a negative result (general IHC practice). |
💡Expected ANXA7 appearanceCall positive when identifiable cells show cytoplasmic staining with possible membranous accentuation, including medium signal in the listed glandular or respiratory epithelial cells; diffuse haze or isolated nuclear staining is unconfirmed (HPA: tissue IHC; general IHC practice).
| Compartment and topology | HPA describes cytoplasmic and membranous staining (HPA: tissue IHC). ANXA7 has no transmembrane segment (UniProt P20073: topology); membrane association should not be read as proof of membrane insertion. |
| Isoforms and tissue context | Isoform 1 is expressed in brain, heart and skeletal muscle; isoform 2 is more abundant in liver, lung, kidney, spleen, fibroblasts and placenta (UniProt P20073: tissue specificity). The supplied IHC record does not identify which isoform an antibody detects. |
| Strength of the tissue evidence | The catalog antibody CAB004312 is IHC Approved, while HPA reports low consistency between antibody staining and RNA data (HPA: antibodies; tissue IHC reliability). Use the listed cell-level observations as reference patterns, with that limitation. |
| Antigen retrieval | No ANXA7-specific retrieval condition or fixation sensitivity is supplied (provided UniProt/HPA records). If optimizing paraffin-section IHC, compare retrieval settings using the same known-positive tissue and controls (general IHC practice). |
| IF/ICC Q: is a localisation pattern validated here? | A: HPA lists no main subcellular location, no cell lines with ICC-IF images and no ICC status for CAB004312 (HPA: subcellular record; antibodies). These data do not establish an IF pattern or supply an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive glandular cells are blank. | The cause cannot be assigned from a blank section; tissue identification, retrieval, primary staining or detection may need checking (general IHC practice). | Confirm the cell type and run a matched positive-control section through the full IHC workflow; compare with HPA’s medium glandular-cell examples (HPA: tissue IHC). |
| Only nuclei show strong colour. | A nuclear-dominant pattern differs from HPA’s cytoplasmic and membranous description (HPA: tissue IHC); its source remains uncertain. | Review morphology and a no-primary control, then repeat under adjusted staining conditions before scoring it as ANXA7 (general IHC practice). |
| Adipocytes or small-intestinal glandular cells look positive. | Those cells are listed as not detected (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible, but neither is established by the image alone. | Compare adjacent expected-positive cells and a no-primary control; address control staining before interpreting the unexpected cells (general IHC practice). |
| Diffuse colour obscures cell boundaries. | Background from blocking, washing or detection can prevent assessment of cellular localisation (general IHC practice). | Inspect a no-primary control, improve the relevant background step and rescore only clearly identifiable cytoplasmic or membranous signal (general IHC practice; HPA: tissue IHC pattern). |
| One positive tissue stains while another does not. | HPA lists cell-specific levels and low consistency with RNA expression; its profile does not promise equal staining across tissues (HPA: tissue IHC). | Compare the same named cell types, specimen quality and matched controls; report each tissue’s observed result instead of imposing one intensity threshold (general IHC practice). |
| IF shows puncta or a compartment unlike the IHC reference. | HPA supplies no main ICC-IF location or cell-line images and gives CAB004312 no ICC status (HPA: subcellular record; antibodies). | Treat the IF localisation as provisional and evaluate it with controls on its separate guide page; use the HPA IHC pattern only as tissue context (HPA: tissue IHC; general IF practice). |
Experiment evidence
IHC & IF images showing ANXA7 staining
PA2076 · IHCIHC analysis of ANXA7 using anti-ANXA7 antibody (PA2076). ANXA7 was detected in a paraffin-embedded section of Human Mammary Cancer tissue.
PA2076 · IFIF analysis of ANXA7 using anti- ANXA7 antibody (PA2076). ANXA7 was detected in immunocytochemical section of A431 cells. Enzyme antigen ret
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138279-ANXA7/tissue/cerebral+cortex#imid_1539032
HPA · Thyroid glandHuman Protein Atlas IHC, Thyroid gland. Source: https://www.proteinatlas.org/ENSG00000138279-ANXA7/tissue/thyroid+gland#imid_1538984
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138279-ANXA7/tissue/colon#imid_1538958
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138279-ANXA7/tissue/liver#imid_1538967
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ANXA7 IHC & IF
🧪Run breast first and score glandular cells for ANXA7 staining (HPA: Medium in breast glandular cells); use adipose tissue as the negative tissue and score adipocytes (HPA: Not detected in adipocytes). On the breast slide, candidate internal negative cells should lack specific DAB staining above background, but their ANXA7-negative status needs independent confirmation because the supplied HPA row reports only glandular cells (HPA: breast glandular cells, Medium).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ANXA7; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control, and ANXA7-knockout tissue as a biological negative (PA2076 caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and biotin before staining breast sections with the caption’s biotin-based, DAB detection system (PA2076 caption: biotinylated secondary and SABC-DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected paraffin-section caption does not state the fixative (PA2076 caption: fixative not stated). The caption uses heat-mediated retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required (PA2076 caption: EDTA retrieval). The supplied evidence does not establish that frozen sections or IF/ICC are easier; biotin-based DAB staining in breast tissue requires attention to endogenous biotin and peroxidase background (PA2076 caption: SABC-DAB; standard IHC practice).
HPA tissue IHC evidence for ANXA7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ANXA7 IHC Tips
Troubleshoot ANXA7 staining by keeping the validated paraffin-section conditions traceable and assessing signal in the relevant cells and compartments.
What retrieval should I try first if ANXA7 staining is weak?
Use heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PA2076). The matched image used 1 μg/ml primary antibody overnight at 4°C after retrieval (caption PA2076). If staining remains weak, compare heating durations on serial sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Check whether stronger signal comes with tissue damage or increased background, using matched control sections (standard IHC practice). Consider another buffer or pH only as a documented fallback; the supplied image gives no comparison between retrieval chemistries (caption PA2076).
Could fixation explain weak or uneven ANXA7 staining?
The PA2076 image identifies a paraffin section but does not report its fixative (caption PA2076). ANXA7-specific sensitivity to fixation is therefore unknown from the supplied evidence (caption PA2076). Record fixative, fixation time, tissue thickness, and processing history before comparing staining across specimens (standard IHC practice). Compare serial sections processed together, using the same pH 8.0 EDTA retrieval and 1 μg/ml antibody starting conditions (datasheet PA2076; caption PA2076). If signal varies with preservation or tissue depth, assess morphology and processing records before changing retrieval or antibody concentration (standard IHC practice).
Where should I expect ANXA7 signal in a paraffin section?
Assess cytoplasmic and membranous staining, the reported broad tissue pattern for ANXA7 (HPA: ubiquitous cytoplasmic and membranous expression). Its calcium and phospholipid binding supports membrane association, while the record lists no transmembrane segment (UniProt P20073 function and topology). UniProt supplies no specific subcellular annotation, so do not require one sharply defined organelle pattern (UniProt P20073 subcellular annotation). Score staining within identifiable cells rather than interpreting isolated DAB deposits as localisation (standard IHC practice). Compare the same cell compartment in reference and experimental sections, and document any predominantly nuclear pattern for further specificity checks (standard IHC practice; HPA: cytoplasmic and membranous profile).
Can tissue differences indicate that my antibody misses an ANXA7 isoform?
ANXA7 has 2 listed isoforms, but the supplied PA2076 image does not map the antibody epitope or establish isoform coverage (UniProt P20073 isoforms; caption PA2076). Isoform 1 is reported in brain, heart, and skeletal muscle, whereas isoform 2 is more abundant in liver, lung, kidney, spleen, fibroblasts, and placenta (UniProt P20073 tissue specificity). Do not assign differential IHC staining to an isoform without epitope mapping or independent isoform-specific evidence (standard IHC interpretation). The record lists acetylation at residue 233, but supplies no evidence that it affects antibody binding (UniProt P20073 modified residues). Compare morphology, retrieval, and controls before proposing epitope masking (standard IHC practice).
How should I relate a multiplex IF result to this IHC stain?
Treat IF as a separate validation exercise: the supplied PA2076 evidence is a paraffin-section chromogenic IHC image, and HPA lists no ICC/IF images (caption PA2076; HPA subcellular). For multiplex IF, pair ANXA7 with a marker identifying the expected glandular cells in breast tissue, then compare cell-level overlap with IHC localisation (HPA: breast glandular cells, medium; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and include single-stain controls when separating channels (standard IF practice). If the mapped epitope is intracellular, optimise permeabilisation for access; ANXA7 has no transmembrane segment, but its epitope side is unspecified here (UniProt P20073 topology; standard IF practice).
How do I distinguish ANXA7 signal from chromogenic background?
The selected PA2076 workflow used 10% goat serum blocking, a biotinylated secondary, streptavidin–biotin detection, and DAB (caption PA2076). Include a no-primary control to reveal secondary or detection-system background on the same tissue (standard IHC practice). Assess endogenous peroxidase and endogenous biotin contributions with appropriate controls before attributing widespread brown deposits to ANXA7 (standard IHC practice; caption PA2076 detection method). Compare staining inside intact cells with signal over folds, damaged edges, pigment, or extracellular debris (standard IHC practice). If background persists, adjust blocking, washing, and detection exposure one variable at a time while retaining a reference section (standard IHC practice).
How should I score ANXA7 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and cytoplasmic or membranous compartment before reviewing treatment groups (HPA: cytoplasmic and membranous profile; standard IHC practice). For breast sections, a percentage of positive glandular cells or an H-score can capture cell-level signal and intensity (HPA: breast glandular cells, medium; standard IHC practice). Report the threshold, intensity categories, sampled area, and whether observers were masked to group assignments (standard IHC practice). If measuring positive-cell density per mm², normalise counts to viable tissue area; for percentage or H-score, use eligible cells as the denominator (standard IHC practice). Exclude necrosis and damaged edges consistently (standard IHC practice).
What makes an ANXA7-positive DAB pattern convincing?
A convincing result places reproducible cytoplasmic or membranous signal within morphologically intact cells (HPA: cytoplasmic and membranous profile; standard IHC practice). In breast tissue, glandular cells provide a reported medium-expression reference, while adipocytes are reported as not detected (HPA: breast glandular cells, medium; HPA: adipocytes, not detected). Investigate predominantly nuclear deposits, staining confined to cut edges or necrosis, and color persisting in a no-primary control (standard IHC practice; HPA: cytoplasmic and membranous profile). Because the selected workflow uses DAB with a biotin-based detection system, evaluate endogenous enzyme and biotin background before calling faint signal positive (caption PA2076; standard IHC practice). Interpret agreement with HPA cautiously because its antibody staining and RNA data have low consistency (HPA: reliability description).
Boster reagents
Best ANXA7 / Annexin A7 IHC Antibodies
PA2076 has IHC data from human mammary cancer paraffin sections and IF data from A431 cells (PA2076 image captions); the catalog lists human reactivity (PA2076 catalog).
Real IHC data
Anti-Annexin VII/ANXA7 Antibody ®
Cat # PA2076
PA2076 is the only SKU with a rendered card and its own IHC figure, showing human mammary cancer paraffin sections (PA2076 IHC caption). Its IF figure shows A431 cells (PA2076 IF caption).
Which to pick: Choose PA2076 for human tissue IHC because its own figure documents paraffin sections, EDTA retrieval at pH 8.0, and 1 μg/ml primary antibody (PA2076 IHC caption); the fixative is unreported (PA2076 IHC caption). For IF/ICC, PA2076 has an A431 cell IF image at 5 μg/ml (PA2076 IF caption). For mouse or rat IHC, consider rabbit monoclonals M04889-1 or M04889-2; both list IHC and human, mouse, and rat reactivity, but neither has an IHC image in the payload (M04889-1 and M04889-2 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P20073 (ANXA7_HUMAN, Annexin A7).
- Human Protein Atlas. ANXA7 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ANXA7 subcellular location (ICC-IF): Highest expression in PC-3: 500.5 nTPM.
- Human Protein Atlas. ANXA7 antibody validation summary (1 antibodies).
- AnnexinA7 promotes epithelial-mesenchymal transition by interacting with Sorcin and contributes to aggressiveness in hepatocellular carcinoma. Cell death & disease 2021 — PMC8556303.
- Tissue microarray analysis delineate potential prognostic role of Annexin A7 in prostate cancer progression. PloS one 2018 — PMC6188866.
- UHRF1 knockdown induces cell cycle arrest and apoptosis in breast cancer cells through the ZBTB16/ANXA7/Cyclin B1 axis. Acta biochimica et biophysica Sinica 2024 — PMC11659777.
- ON-bipolar cell gene expression during retinal degeneration: Implications for optogenetic visual restoration. Experimental eye research 2021 — PMC8214074.
- PubMed PMID:2542947 — UniProt-cited evidence.
- PubMed PMID:7515686 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ANXA7 (Annexin A7). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PA2076). Validated IHC antibody: PA2076 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human Mammary Cancer tissue; fixative not specified (datasheet PA2076); 2. Fixation: Image fixative and duration unreported (datasheet PA2076); 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 PA2076); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PA2076); 8. Primary antibody: Rabbit anti-ANXA7, 0.5-1μg/ml (datasheet PA2076); 9. Primary incubation: Overnight at 4 °C (datasheet PA2076). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA levels show low consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Isoform abundance differs by tissue (UniProt). Evidence: UniProt P20073, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in PC-3: 500.5 nTPM, Datasheet PA2076, PMC6188866, PMC11659777. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANXA7/immunohistochemistry-ihc-guide.