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AAMP / Angio-associated migratory cell protein · IHC design guide
Design Immunohistochemistry for AAMP
Plan chromogenic AAMP IHC in paraffin sections using the catalog antibody at 2–5 μg/mL (datasheet PB9123). Assess cytoplasmic staining and use smooth muscle as a tissue reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AAMP 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 in tissue (HPA tissue IHC); membrane also annotated (UniProt) | |
| Staining pattern | Cytoplasmic staining, most abundant in smooth muscle (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9123) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Skeletal muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across specimens. (selected-SKU IHC image PB9123) | |
| Caveat | Smooth muscle content can shift apparent tissue signal (HPA tissue IHC) | |
| Regulation | Expression in activated T cells is reported (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one 1–434 chain (UniProt) |
Section 1
Recommended AAMP IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet PB9123) with the published glioma IHC protocol (PMC9341442).
| Sample | Paraffin-embedded human testis cancer tissue; fixative not specified (datasheet PB9123) |
| Fixation | Image fixative and duration unreported (datasheet PB9123); 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 PB9123); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9123) |
| Primary antibody | Rabbit anti-AAMP, 2-5μg/ml (datasheet PB9123) |
| Primary incubation | Overnight at 4 °C (datasheet PB9123) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9123) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AAMP-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, most abundant in smooth muscle. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet PB9123). To reproduce the published glioma staining, use its cell conditioner pretreatment (PMC9341442).
Section 2
What Is the Expected AAMP Staining Pattern?
AAMP staining in paraffin sections is expected mainly in the cytoplasm, with strong signal in smooth muscle cells and several other cell populations (HPA: tissue IHC, Approved; High in smooth muscle). Membrane-associated staining can also fit its annotation, although AAMP has no annotated transmembrane segment (UniProt Q13685: cell membrane, cytoplasm; topology). Interpret intensity by cell type: HPA reports low tissue specificity and only medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Strong cytoplasmic staining in smooth muscle cells, with cellular outlines still discernible. | This matches the most abundant compartment and cell type in the tissue profile (HPA: general cytoplasmic expression; High in smooth muscle cells). Use the distribution across cells as well as intensity when judging a positive section. |
| Cytoplasmic signal in adipocytes or alveolar cells, or signal in the named ciliated cell populations. | These can be expected positives (HPA: High in adipocytes, alveolar cells, and specified ciliated cells). Cilia-associated staining is documented in fallopian tube and nasopharynx; do not require every listed cell type to show the same pattern. |
| Predominantly nuclear staining, without a convincing cytoplasmic or membrane-associated pattern. | This does not match the reported IHC profile or UniProt locations (HPA: general cytoplasmic expression; UniProt Q13685: cytoplasm, cell membrane). Consider nonspecific staining or a detection artefact before assigning it to AAMP. |
| Strong staining of skeletal myocytes, particularly when adjacent expected positive cells are weak. | Treat this as discordant with the cell-level reference, rather than proof of cross-reactivity (HPA: skeletal muscle myocytes Not detected). UniProt reports protein expression in skeletal muscle, so compare the exact stained cell population and antibody controls. |
| Diffuse color across cells, stroma, and blank areas, or no staining in a suitable positive section. | A pattern that ignores cell boundaries is hard to score as AAMP; absent signal in smooth muscle cells conflicts with a reported high-staining reference (HPA: general cytoplasmic expression; High in smooth muscle). Either finding needs control review. |
💡Expected AAMP appearanceCall a section positive when identifiable smooth muscle cells show strong, mainly cytoplasmic staining, allowing membrane-associated signal; nuclear-only color, diffuse deposit, or strong skeletal-myocyte staining is suspect and should be checked against controls (HPA: tissue IHC profile; UniProt Q13685: subcellular location).
| Cell population and tissue choice | Smooth muscle cells provide a strong reference, while adipocytes, alveolar cells, ovarian stromal cells, and named ciliated populations are also reported High (HPA: tissue IHC). Sample and score the specified cells; whole-tissue intensity can obscure cell-level differences. |
| Reference-negative interpretation | Skeletal myocytes are reported Not detected by HPA, but UniProt lists skeletal muscle among tissues with protein expression (HPA: tissue IHC; UniProt Q13685: tissue specificity). A negative myocyte result is a useful comparison, not proof that the whole tissue lacks AAMP. |
| Compartment and protein annotation | The cytoplasmic tissue profile agrees with UniProt's cytoplasm annotation; membrane-associated staining is also plausible (HPA: tissue IHC; UniProt Q13685: cell membrane, cytoplasm). No transmembrane segment is annotated, so do not demand a sharp membrane-only rim (UniProt Q13685: topology). |
| IHC evidence strength | The tissue profile is Approved, with medium consistency between staining and RNA data; the listed IHC antibody CAB017717 is Approved (HPA: tissue IHC reliability; antibody validation). These labels support use as a reference but do not establish specificity for every cell or slide condition. |
| Processing and epitope limits | UniProt lists one 1–434 chain, no signal peptide or propeptide, no annotated glycosylation sites, and no isoforms (UniProt Q13685: processing, glycosylation, isoforms). The supplied sources give no epitope position or AAMP-specific fixation sensitivity; neither can explain a failed section here. |
| IF/ICC Q: Should its pattern match chromogenic tissue IHC? | A: Judge IF/ICC on its own page. HPA reports mainly microtubules, plus plasma membrane, cytosol, cytokinetic bridge, and primary cilium locations in ICC-IF (HPA: subcellular). Those observations do not require every paraffin tissue section to resolve those structures. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a section expected to contain smooth muscle cells. | The result conflicts with a High cell-level reference, but the slide alone cannot identify the failed step (HPA: smooth muscle cells High). | Confirm that smooth muscle cells are present and identifiable; review the positive control, antibody dilution, retrieval, detection reagents, and counterstain using the applicable IHC workflow. Treat retrieval checks as general FFPE practice, not an AAMP-specific fixation claim. |
| Strong nuclear-only color. | The dominant compartment is discordant with the reported cytoplasmic tissue pattern and UniProt locations (HPA: tissue IHC; UniProt Q13685: subcellular location). | Inspect the no-primary control and detection background, then compare with a section containing a known positive cell population (HPA: smooth muscle cells High). Do not score nuclear-only staining as confirmed AAMP. |
| Strong color in skeletal myocytes. | HPA reports skeletal myocytes as Not detected, although UniProt reports expression in skeletal muscle without assigning it to myocytes (HPA: tissue IHC; UniProt Q13685: tissue specificity). | Verify myocyte identity and compare staining with neighboring cells and a no-primary control. If strong myocyte signal persists, record the discordance and check antibody specificity before scoring it as AAMP. |
| Color appears throughout cells, stroma, or empty areas. | Diffuse deposit lacks the cell-linked cytoplasmic distribution used for interpretation; nonspecific binding or endogenous detection activity are general IHC possibilities (HPA: tissue IHC profile; standard IHC practice). | Compare with no-primary and detection controls; review blocking, wash steps, chromogen development, and endogenous enzyme blocking where relevant. Score only interpretable cellular staining after background is controlled. |
| Patchy staining within an otherwise suitable section. | Variation in section handling or reagent coverage is a general IHC possibility; the supplied sources do not establish AAMP-specific fixation sensitivity (standard IHC practice; HPA: tissue IHC scope). | Check whether positive cells occur in the weak area, then inspect section integrity, reagent coverage, and the positive control. Repeat the stain if these checks leave the local absence uninterpretable. |
| Weak staining is seen in a tissue outside the listed High populations. | AAMP has low tissue specificity, and HPA records several Low cell-level results; intensity alone does not define a new positive or negative tissue class (HPA: RNA specificity; tissue IHC Low entries). | Identify and score the stained cell type, compare its compartment with the cytoplasmic reference, and document the result alongside controls (HPA: tissue IHC profile). Avoid extending the reference to an unlisted cell population without validation. |
Experiment evidence
IHC & IF images showing AAMP staining
PB9123 · IHCIHC analysis of AAMP using anti-AAMP antibody (PB9123). AAMP was detected in a paraffin-embedded section of human testis cancer tissue. Heat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000127837-AAMP/tissue/cerebral+cortex#imid_4517492
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000127837-AAMP/tissue/small+intestine#imid_4517401
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000127837-AAMP/tissue/colon#imid_4517426
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000127837-AAMP/tissue/liver#imid_4517411
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017717; green = AAMP, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017717; green = AAMP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AAMP IHC & IF
🧪Run adipose tissue first and expect staining in adipocytes (HPA: High in adipocytes). Run skeletal muscle as the negative tissue and expect myocytes to lack detectable staining (HPA: Not detected in myocytes); on the adipose slide, assess non-adipocyte cells individually as internal background references rather than assuming all are negative.
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Skeletal muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AAMP in A-431, U-251MG, U2OS, MCF-7, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), NIH 3T3, with annotated localisation: Microtubules (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (PB9123 caption: rabbit primary antibody). Use AAMP knockout material or a validated peptide-blocking control if available; check for endogenous peroxidase signal in chromogenic sections and lipid-related autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9123 paraffin-section caption does not state a fixative (PB9123 caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required (PB9123 caption: EDTA retrieval); the supplied evidence does not establish whether frozen sections or IF are easier. In adipose tissue, lipid extraction can leave empty vacuoles that complicate cell identification (standard IHC practice).
HPA tissue IHC evidence for AAMP
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AAMP IHC Tips
Troubleshoot AAMP staining in paraffin sections by checking retrieval, cell identity, subcellular pattern and detection controls (datasheet PB9123; HPA tissue IHC; UniProt Q13685).
What retrieval should I use when AAMP staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet PB9123). The documented paraffin-section stain used 2 μg/ml primary antibody overnight at 4°C, so repeat that combination with a positive control before changing retrieval (datasheet PB9123). Compare sections processed together and check whether weak signal affects expected cytoplasmic staining in smooth muscle as well as the specimen of interest (HPA tissue IHC: general cytoplasmic expression, most abundant in smooth muscle; standard IHC practice). If staining remains weak, test a different retrieval condition on adjacent sections while holding antibody concentration and chromogen development constant, and assess tissue damage alongside signal (standard IHC practice).
How should I troubleshoot possible fixation effects on AAMP IHC?
AAMP-specific sensitivity to fixation is unknown from the supplied evidence; the selected image documents a paraffin-embedded section but does not report its fixative (datasheet PB9123). Record the specimen’s actual fixative and processing history, then compare sections with known processing histories using the same EDTA pH 8.0 retrieval and staining run (datasheet PB9123; standard IHC practice). Inspect morphology and assess whether loss of signal is widespread or confined to poorly preserved areas before adjusting staining conditions (standard IHC practice). Use an expected positive cell population as a run control, while treating differences between fixation groups as observations requiring validation rather than an established AAMP-specific effect (HPA tissue IHC: high in smooth muscle cells; standard IHC practice).
Which AAMP staining compartments should I expect in tissue sections?
Assess cytoplasmic staining first: tissue IHC reports general cytoplasmic expression, most abundant in smooth muscle (HPA tissue IHC). Membrane-associated signal can be biologically plausible because AAMP is assigned to the cell membrane and cytoplasm, although it has no annotated transmembrane segment (UniProt Q13685: subcellular location and topology). Cell imaging also places AAMP mainly at microtubules, with additional plasma membrane, cytosol and ciliary locations; those finer patterns may be difficult to resolve with chromogenic tissue IHC (HPA subcellular; standard IHC practice). Judge distribution against cell morphology and a counterstain, and investigate predominantly nuclear or diffuse extracellular staining before calling it specific (standard IHC practice).
Could an isoform or epitope issue explain variable AAMP staining?
The supplied AAMP record lists 0 isoforms and one chain spanning residues 1–434, so an annotated isoform difference does not explain variable staining here (UniProt Q13685: isoforms and processing). The antibody’s epitope position is not supplied, preventing a prediction about which part of AAMP retrieval might expose (datasheet PB9123: epitope unspecified). AAMP has an annotated phosphoserine at residue 20, no annotated glycosylation sites and no transmembrane segment, but none establishes how this antibody binds in tissue (UniProt Q13685: modified residues, glycosylation and topology). Compare staining across identically processed sections and verify discordant patterns with an independently validated antibody if available (standard IHC practice).
How can IF help resolve ambiguous AAMP localisation seen by IHC?
For a separate IF experiment, multiplex AAMP with a marker identifying the suspected cell population, such as smooth muscle, and compare the two signals cell by cell (HPA tissue IHC: high in smooth muscle cells; standard IF practice). Choose spectrally separated fluorophores and include unstained tissue to assess autofluorescence before assigning faint signal to AAMP (standard IF practice). Because AAMP has reported cytosolic and microtubule localisation as well as plasma membrane localisation, select permeabilisation according to the antibody epitope’s accessibility and test it against a matched condition (HPA subcellular; standard IF practice). The epitope side is unspecified, so do not infer its accessibility from AAMP’s lack of a transmembrane segment alone (datasheet PB9123: epitope unspecified; UniProt Q13685: topology).
What should I check when AAMP DAB staining has high background?
Compare a section processed without primary antibody with the test section to identify signal arising from secondary detection or endogenous tissue activity (standard IHC practice). Include an appropriate peroxidase block and inspect DAB development time; the documented paraffin-section workflow used a peroxidase-conjugated secondary and DAB (standard IHC practice; datasheet PB9123). The image workflow also used 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C, which provide a documented starting condition for this antibody (datasheet PB9123). If background persists, adjust blocking, primary concentration or development one variable at a time while retaining a positive-control section (standard IHC practice).
How should I quantify AAMP staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; cytoplasmic AAMP expression is reported across tissues and is most abundant in smooth muscle (HPA tissue IHC). For a cell-based comparison, record the percentage of positive cells and staining intensity, then calculate an H-score using one consistent threshold and sampling rule (standard IHC practice). For spatial questions, report positive-cell density per mm² within a defined tissue area, excluding folds, necrosis and section edges by a prespecified rule (standard IHC practice). Normalise counts to eligible cells or evaluable tissue area, and report smooth muscle-rich regions separately when their abundance differs between samples (HPA tissue IHC; standard IHC practice).
How do I distinguish credible AAMP staining from artefact?
A convincing result should follow identifiable cells and a plausible cytoplasmic pattern, especially in smooth muscle, where tissue IHC reports high staining (HPA tissue IHC). Membrane-associated signal can also fit the protein record, but predominantly nuclear staining lacks support from the supplied localisation evidence and warrants validation (UniProt Q13685: cell membrane and cytoplasm; HPA subcellular). Treat staining concentrated at section edges, folds or necrotic areas as suspect, and use a no-primary control to investigate endogenous peroxidase or detection background (standard IHC practice). Interpret weak or discordant tissue staining cautiously because the reported HPA tissue-IHC reliability is Approved with medium consistency between staining and RNA expression (HPA tissue IHC).
Boster reagents
Best AAMP / Angio-associated migratory cell protein IHC Antibodies
PB9123 has a human paraffin-section IHC figure (catalog image caption); M07409 lists IF/ICC and Human, Mouse, Rat reactivity, with no supplied IF figure (catalog applications/reactivity/images).
Real IHC data
Anti-AAMP Antibody ®
Cat # PB9123
PB9123 will render with its IHC figure from a paraffin-embedded human testis cancer section (catalog image caption). M07409 lists IF/ICC and Human, Mouse, Rat reactivity, but has no supplied IF figure (catalog applications/reactivity/images).
Which to pick: Choose PB9123 for tissue IHC: its own image caption documents EDTA pH 8.0 retrieval and 2 μg/ml antibody on a human paraffin section; the fixative is unreported (PB9123 image caption). Choose M07409 for IF/ICC because those applications and a 1:50–1:200 dilution are listed, although no IF image is supplied (M07409 catalog applications/dilution/images). Both list Human, Mouse and Rat reactivity for cross-species planning, but PB9123’s stated paraffin-section IHC dilution is for Human (catalog reactivity/dilution).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13685 (AAMP_HUMAN, Angio-associated migratory cell protein).
- Human Protein Atlas. AAMP tissue IHC expression (reliability: Approved).
- Human Protein Atlas. AAMP subcellular location (ICC-IF): Mainly localized to the microtubules. In addition localized to the plasma membrane, cytosol, cytokinetic bridge, primary cilium and primary cilium tip..
- Human Protein Atlas. AAMP antibody validation summary (2 antibodies).
- AAMP promotes colorectal cancermetastasis by suppressing SMURF2-mediatedubiquitination and degradation of RhoA. Molecular therapy oncolytics 2021 — PMC8633529.
- Pan-cancer analysis from multi-omics data reveals AAMP as an unfavourable prognostic marker. European journal of medical research 2023 — PMC10373365.
- AAMP is a binding partner of costimulatory human B7-H3. Neuro-oncology advances 2022 — PMC9341442.
- PubMed PMID:7743515 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:10329261 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AAMP (Angio-associated migratory cell protein). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); membrane also annotated (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9123). Validated IHC antibody: PB9123 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Skeletal muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human testis cancer tissue; fixative not specified (datasheet PB9123); 2. Fixation: Image fixative and duration unreported (datasheet PB9123); 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 PB9123); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9123); 8. Primary antibody: Rabbit anti-AAMP, 2-5μg/ml (datasheet PB9123); 9. Primary incubation: Overnight at 4 °C (datasheet PB9123). Watch for: Fixation: Keep paraffin-section fixation consistent across specimens. (selected-SKU IHC image PB9123); Caveat: Smooth muscle content can shift apparent tissue signal (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; one 1–434 chain (UniProt); Regulation: Expression in activated T cells is reported (UniProt). Evidence: UniProt Q13685, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the microtubules. In addition localized to the…, Datasheet PB9123, PMC9341442. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AAMP/immunohistochemistry-ihc-guide.