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ANXA7 / Annexin A7 · IHC design guide
Design Immunohistochemistry for ANXA7
Plan ANXA7 staining in paraffin sections using its reported cytoplasmic and membranous tissue pattern (HPA tissue IHC). This guide covers controls, interpretation, and the documented 2 isoforms (UniProt).
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 | Widespread cytoplasmic and membranous signal (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PA2076) | |
| Caveat | Antibody staining has low agreement with RNA data (HPA tissue IHC) | |
| Regulation | Isoform abundance varies by tissue (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended ANXA7 IHC & IF Protocols
The catalog antibody IHC-P protocol (datasheet: PA2076) sits alongside published ANXA7 staining of prostate cancer tissue microarrays (PMC6188866) and tumor tissue sections (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 retrieval (datasheet: PA2076); for optimization, compare the published microwave sodium citrate method (PMC11659777).
Section 2
What Is the Expected ANXA7 Staining Pattern?
Expect mainly cytoplasmic and membranous ANXA7 staining in paraffin sections, including medium staining in several glandular, respiratory epithelial, lymphoid and hematopoietic cell populations (HPA tissue IHC). ANXA7 has no transmembrane segment, so membranous staining describes an observed pattern, not a membrane-spanning protein (UniProt P20073 topology). HPA rates the tissue IHC profile Approved, while reporting low consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining, with or without a membranous component, in glandular or respiratory epithelial cells. | This fits the reported pattern. HPA records medium staining in glandular cells of adrenal gland, breast, cervix, endometrium and epididymis, and in bronchial respiratory epithelial cells (HPA tissue IHC). Compare cells within the section: the profile is described as ubiquitous, so a single stained cell population is not required in every tissue (HPA tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic or membranous signal. | This differs from the reported tissue IHC pattern and warrants a specificity check (HPA tissue IHC). Inspect the negative control and repeat with an appropriate control tissue before interpreting nuclear signal as ANXA7. UniProt supplies no subcellular localisation annotation, so the tissue IHC pattern is the relevant reference here (UniProt P20073; HPA tissue IHC). |
| Strong signal in adipocytes or small-intestinal glandular cells. | HPA reports ANXA7 as not detected in those respective cell populations (HPA tissue IHC). Check whether the signal follows the cells rather than pigment, section edges or endogenous detection activity (general IHC practice). Persistent staining could reflect cross-reactivity or assay background; the HPA observations are comparison points, not absolute biological negatives. |
| Diffuse colour covers stroma, empty spaces or many unrelated cell types. | A widespread haze obscures the cytoplasmic and membranous cellular pattern reported by HPA (HPA tissue IHC). Background can arise from nonspecific reagent binding or endogenous detection activity (general IHC practice). Judge localisation only after examining a negative control and areas where tissue structure remains clear. |
| No staining in a section containing HPA-reported medium-positive cells. | A blank result in, for example, breast glandular cells or bronchial respiratory epithelial cells conflicts with the reported medium staining (HPA tissue IHC). First confirm that the expected cells are present and that the detection run worked. Then review the antibody dilution and retrieval procedure against the chosen IHC protocol (general IHC practice). |
💡Expected ANXA7 appearanceCall a result positive when cytoplasmic, often membranous, signal is discernible in the relevant cells, with medium staining in HPA-listed positive populations; dominant nuclear staining, diffuse haze or strong signal in HPA-listed not-detected cells merits investigation (HPA tissue IHC).
| Tissue and cell context | HPA reports medium staining in selected glandular, respiratory epithelial, lymphoid and hematopoietic cells, but no detection in adipocytes or small-intestinal glandular cells (HPA tissue IHC). Score the named cell population rather than treating the whole organ as uniformly positive or negative. |
| Isoform context | UniProt lists two isoforms: 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). No supplied antibody epitope or isoform-selectivity data establish different IHC staining for the two isoforms. |
| Membrane interpretation | ANXA7 binds calcium and phospholipids and promotes membrane fusion (UniProt P20073 function). HPA describes cytoplasmic and membranous staining (HPA tissue IHC). Because UniProt reports no transmembrane segment, a membrane-associated appearance should not be scored as evidence of a membrane-spanning topology (UniProt P20073 topology). |
| Evidence strength | The listed antibody, CAB004312, has Approved IHC status, while HPA notes low consistency between staining and RNA expression (HPA antibodies; HPA tissue IHC). Treat the profile as a guide for comparison and investigate unexpected cells or compartments with controls before making a biological claim. |
| IF/ICC: what pattern is established? | HPA provides no main subcellular location or cell-line ICC-IF images for ANXA7 (HPA subcellular). The tissue IHC pattern can inform a localisation question, but this record does not establish an IF/ICC pattern or an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells remain unstained. | The sampled section may lack the specified cells, or the IHC run may have failed. | Verify cell identity and include a section with HPA-reported medium staining, such as breast glandular cells; check run controls and the chosen IHC protocol (HPA tissue IHC; general IHC practice). |
| Only nuclear staining is visible. | The distribution does not match HPA's cytoplasmic and membranous tissue profile (HPA tissue IHC). | Compare with a negative control and an HPA-listed positive cell population; withhold a nuclear localisation claim unless independently supported (general IHC practice). |
| Adipocytes or small-intestinal glandular cells stain strongly. | Those cells are listed as not detected, so cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Check cell identity, inspect the negative control, and assess whether signal tracks tissue structures or nonspecific deposits (general IHC practice). |
| Diffuse background makes cell borders hard to judge. | Nonspecific binding or endogenous detection activity may obscure localisation (general IHC practice). | Review blocking, washes and detection controls; score ANXA7 only where cellular morphology and the reported cytoplasmic or membranous pattern remain clear (general IHC practice; HPA tissue IHC). |
| A section looks weaker than another HPA-listed tissue. | HPA lists staining by cell population and reports low staining–RNA consistency; an organ-wide comparison can mislead (HPA tissue IHC). | Compare like cell populations and record intensity and compartment separately. Do not infer isoform abundance from staining alone; antibody isoform selectivity is unspecified (UniProt P20073; HPA tissue IHC). |
| Membranous colour is interpreted as exclusive cell-surface expression. | HPA also reports cytoplasmic staining, and ANXA7 lacks a transmembrane segment (HPA tissue IHC; UniProt P20073 topology). | Document both compartments as observed and avoid assigning a membrane-spanning localisation from chromogenic IHC alone (HPA tissue IHC; UniProt P20073 topology). |
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 expect glandular cells to stain (HPA: Medium in breast glandular cells); run adipose tissue as the negative and expect adipocytes to lack detectable staining (HPA: Not detected in adipocytes). On the breast slide, use cells without specific signal as internal background references, without assuming a particular cell type is ANXA7-negative.
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 matched rabbit IgG isotype control (caption: rabbit primary antibody), and ANXA7-knockout material as a biological negative. Quench endogenous peroxidase and check for endogenous biotin when using the caption’s biotin–avidin/DAB detection scheme (selected PA2076 IHC caption).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected PA2076 paraffin-section caption does not state a fixative. The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected PA2076 IHC caption). There is no supplied comparison showing that frozen sections or IF are easier; with the caption’s biotin–avidin detection, endogenous biotin can contribute background in breast tissue (selected PA2076 IHC caption).
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 in paraffin sections by checking retrieval, tissue processing, compartment pattern and controls before interpreting chromogenic signal.
What should I change when ANXA7 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PA2076). The selected tissue image used that retrieval, followed by 1 μg/ml primary antibody overnight at 4°C, so match those documented conditions before changing several variables at once (caption PA2076). If staining remains weak, compare a second retrieval condition on adjacent sections while keeping detection and imaging constant; use tissue morphology and an established positive control to judge whether heating was excessive (standard IHC practice). Include a no-primary control because stronger retrieval can also expose background staining (standard IHC practice).
Could fixation explain weak or uneven ANXA7 staining?
Target-specific fixation sensitivity is unknown: the selected ANXA7 image identifies a paraffin section but does not state its fixative (caption PA2076). Record the actual fixative and fixation time for each specimen, then compare sections processed alike before attributing a difference to ANXA7 abundance (standard IHC practice). Keep retrieval at EDTA pH 8.0 and primary antibody at 1 μg/ml during that comparison, since those conditions are documented for the selected antibody (caption PA2076). Check preservation, tissue edges and control sections; neither the HPA staining profile nor ANXA7 topology establishes a fixation effect (HPA tissue IHC; UniProt P20073 topology).
Should ANXA7 stain cytoplasm, membranes or both in tissue sections?
Assess cytoplasmic and membranous signal together, since the tissue atlas describes ANXA7 expression in both compartments (HPA tissue IHC). ANXA7 binds calcium and phospholipids and participates in membrane fusion, while its sequence has no transmembrane segment; a membrane-associated pattern therefore does not require integral membrane insertion (UniProt P20073 function and topology). Compare glandular cells with adjacent structures within the same section: breast glandular cells show medium staining in the atlas, whereas adipocytes are listed as not detected (HPA tissue IHC). Treat an exclusively nuclear or diffuse acellular deposit as suspect until controls and section quality support it (HPA tissue IHC; standard IHC practice).
Can this stain distinguish ANXA7 isoforms or reveal epitope masking?
Do not assign the stain to a particular isoform without an epitope map or isoform-selective validation; ANXA7 has 2 reported isoforms (UniProt P20073 isoforms). Isoform 1 is reported in brain, heart and skeletal muscle, while isoform 2 is more abundant in liver, lung, kidney, spleen, fibroblasts and placenta (UniProt P20073 tissue specificity). The record lists an acetylated lysine at residue 233, but supplies no evidence that this modification changes antibody binding (UniProt P20073 modified residues). If staining differs across specimens, check antibody epitope documentation and compare matched controls under the documented EDTA pH 8.0 retrieval conditions before proposing an isoform explanation (datasheet PA2076; standard IHC practice).
How should I check an ANXA7 pattern with immunofluorescence?
For a separate IF assay, pair ANXA7 with a marker for the expected cell type and compare whether their signals occupy the same cells; breast glandular cells are a documented tissue example (HPA tissue IHC; standard IF practice). Choose fluorophores and imaging channels after measuring tissue autofluorescence, and include single-stain controls when multiplexing (standard IF practice). Because ANXA7 lacks a transmembrane segment and tissue staining includes cytoplasm and membranes, use a permeabilisation condition that admits antibody to cytoplasmic epitopes while preserving membrane-associated signal (UniProt P20073 topology; HPA tissue IHC; standard IF practice). The supplied subcellular atlas has no ICC/IF images, so verify the fluorescent pattern independently rather than treating the chromogenic image as IF validation (HPA subcellular; caption PA2076).
How can I reduce diffuse or misleading ANXA7 background?
Begin with a no-primary control and inspect whether deposits follow tissue edges, damaged areas or the detection chemistry rather than cells (standard IHC practice). The selected protocol blocked with 10% goat serum, used 1 μg/ml primary overnight at 4°C, and detected signal through a biotin-based system with DAB (caption PA2076). If background persists, titrate the primary and detection reagents separately, check endogenous biotin when using that detection system, and include a peroxidase block as a general chromogenic IHC step (standard IHC practice). Judge changes against a matched positive section because widespread cytoplasmic and membranous staining is part of the reported ANXA7 tissue profile (HPA tissue IHC).
What is a defensible way to quantify ANXA7 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and an intensity-based H-score using the same thresholds across specimens (standard IHC practice). If counting positive cells per mm², normalise to the area of viable tissue or the relevant cell compartment, and exclude folds, necrosis and blank spaces (standard IHC practice). Record cytoplasmic and membranous scores separately because both appear in the atlas profile (HPA tissue IHC). Use matched processing, exposure and controls, and interpret small differences cautiously because the atlas marks tissue staining as Approved while noting low consistency with RNA expression (HPA tissue IHC).
When should an apparent ANXA7-positive cell be considered an artefact?
A convincing result follows cell boundaries or cytoplasm in viable tissue and agrees with matched controls; the reported ANXA7 profile is cytoplasmic and membranous (HPA tissue IHC; standard IHC practice). Check cell identity before calling a region positive: breast glandular cells have medium atlas staining, while adipocytes are listed as not detected (HPA tissue IHC). Be skeptical of isolated nuclear signal, edge-only staining, necrotic deposits or pigment that remains in a no-primary control (HPA tissue IHC; standard IHC practice). In DAB sections, verify that endogenous peroxidase and the biotin-based detection system are controlled before interpreting brown signal as ANXA7 (caption PA2076; standard IHC practice).
Boster reagents
Best ANXA7 / Annexin A7 IHC Antibodies
PA2076 has IHC data from a human mammary cancer paraffin section and IF data from A431 cells (PA2076 image captions). Two additional antibodies list human, mouse and rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-Annexin VII/ANXA7 Antibody ®
Cat # PA2076
PA2076 is the card shown: its IHC image shows staining in a human mammary cancer paraffin section (PA2076 IHC caption). Its IF image shows staining in A431 cells (PA2076 IF caption).
Which to pick: Choose PA2076 for human paraffin-section IHC: its image documents EDTA retrieval at pH 8.0 and primary antibody at 1 μg/ml; the fixative is unreported (PA2076 IHC caption). For IF/ICC, PA2076 has an A431-cell IF image, while M04889-1 lists IF/ICC without an image in this payload (PA2076 IF caption; M04889-1 catalog applications and image alts). For cross-species IHC, M04889-1 and M04889-2 are rabbit monoclonals listing human, mouse and rat reactivity, but neither has an IHC image here; only M04889-1 also lists IF/ICC (catalog clones, reactivity, applications and image alts).
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: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PA2076); Caveat: Antibody staining has low agreement with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Isoform abundance varies 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.