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AAMP / Angio-associated migratory cell protein · IHC design guide
Design Immunohistochemistry for AAMP
Plan AAMP chromogenic IHC on paraffin sections using the catalog antibody’s validated staining conditions (datasheet PB9123). Compare cytoplasmic staining across cells, with smooth muscle as a strong reference and skeletal myocytes as a useful contrast (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 localisation annotated (UniProt) | |
| Staining pattern | Cytoplasmic staining, most abundant in smooth muscle cells (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 fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Skeletal myocytes show no detected staining despite reported expression (HPA tissue IHC; UniProt) | |
| Regulation | Abundance varies by cell type (HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms or cleavage; epitope sidedness unknown (UniProt) |
Section 1
Recommended AAMP IHC & IF Protocols
The catalog antibody’s IHC protocol (datasheet PB9123) is accompanied by a published human 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 EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet PB9123). The published protocol reports Cell Conditioner pretreatment without a pH or heating condition (PMC9341442).
Section 2
What Is the Expected AAMP Staining Pattern?
AAMP is reported at the cell membrane and in the cytoplasm, with no transmembrane segment annotated (UniProt Q13685). In paraffin section IHC, expect chiefly cytoplasmic staining, especially in smooth muscle cells, with strong staining also reported in adipocytes and several other cell types (HPA tissue IHC). HPA rates its tissue IHC evidence Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Strong cytoplasmic staining in smooth muscle cells, with cellular detail retained. | This fits the most abundant tissue pattern reported for AAMP (HPA tissue IHC: High in smooth muscle cells). Compare the stained cells with tissue morphology before scoring the section (general IHC practice). |
| Predominantly nuclear staining, or a pattern confined to an unexpected compartment. | An isolated nuclear pattern does not match the reported cytoplasmic tissue pattern or the annotated cytoplasm and cell membrane locations (HPA tissue IHC; UniProt Q13685). Check controls and staining conditions before assigning it to AAMP (general IHC practice). |
| Strong staining in skeletal myocytes, while expected positive cells are weak or absent. | HPA reports AAMP as not detected in skeletal myocytes (HPA tissue IHC). Consider cross-reactivity or detection artefact, and compare with a known positive tissue and negative reagent control (general IHC practice). |
| Uniform colour across tissue and empty spaces, obscuring cell boundaries. | A cell independent haze cannot establish AAMP localisation (general IHC practice). Assess blocking, washing and detection background with a negative reagent control before interpreting intensity (general IHC practice). |
| No staining in a section containing recognizable smooth muscle cells. | This conflicts with HPA's High smooth muscle cell staining (HPA tissue IHC), but does not by itself identify the failed step. Review tissue preservation, retrieval, antibody dilution and detection controls as general IHC checks (general IHC practice). |
💡Expected AAMP appearanceCall a result positive when recognizable smooth muscle cells show strong, chiefly cytoplasmic staining (HPA tissue IHC: High); isolated nuclear colour or staining in empty spaces is suspect (UniProt Q13685; general IHC practice).
| Tissue and cell selection | Smooth muscle cells offer a reported High reference, whereas skeletal myocytes are reported Not detected (HPA tissue IHC). Low staining in selected epithelia and cardiomyocytes makes those cells less clear as positive references (HPA tissue IHC). |
| Compartment interpretation | UniProt places AAMP at the cell membrane and in the cytoplasm and annotates no transmembrane segment (UniProt Q13685). HPA tissue IHC describes general cytoplasmic expression; assess membrane colour against cell morphology and controls (HPA tissue IHC; general IHC practice). |
| Strength of tissue evidence | The tissue profile is Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC). One listed antibody has Approved IHC status; the other has no listed IHC status (HPA antibodies). Avoid treating either designation as proof of every stained cell (general IHC practice). |
| IF/ICC Q&A: should the pattern look identical? | IF/ICC images mainly localize AAMP to microtubules, with additional plasma membrane, cytosol, cytokinetic bridge and cilium locations (HPA subcellular ICC-IF). Those cell images inform localisation but do not establish that every feature will resolve in chromogenic paraffin sections (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Smooth muscle cells show no signal. | A missing signal in a reported High cell type may reflect an unsuccessful staining run (HPA tissue IHC; general IHC practice). | Check tissue morphology, the run's positive control, retrieval, catalog antibody dilution and chromogenic detection in sequence (general IHC practice). No AAMP specific retrieval condition is supplied. |
| Only nuclei are strongly coloured. | The compartment conflicts with reported cytoplasmic tissue staining and UniProt localisation (HPA tissue IHC; UniProt Q13685). | Review counterstain and detection controls, then repeat with controlled staining conditions before scoring nuclei as positive (general IHC practice). |
| Skeletal myocytes stain strongly. | That contrasts with the HPA Not detected observation in skeletal myocytes (HPA tissue IHC); cross-reactivity or nonspecific detection is possible (general IHC practice). | Compare a known positive cell population and a negative reagent control in the same run; interpret any residual myocyte colour cautiously (general IHC practice). |
| Broad brown haze obscures cells. | Background from detection chemistry or inadequate washing can conceal cellular staining (general IHC practice). | Inspect a negative reagent control, check blocking and washing, and assess endogenous detection activity as appropriate to the chromogen system (general IHC practice). |
| Staining appears strong only in an unexpected tissue cell population. | AAMP has broad tissue expression, but HPA assigns different levels to specific cell types (HPA tissue IHC). Morphology or nonspecific staining may account for the discrepancy (general IHC practice). | Identify cells morphologically and compare their reported HPA level; use controls before calling an unexpected population AAMP positive (HPA tissue IHC; general IHC practice). |
| IF/ICC shows fine structures absent from the IHC section. | HPA ICC-IF reports microtubules and additional fine structures, while tissue IHC describes a general cytoplasmic pattern (HPA subcellular ICC-IF; HPA tissue IHC). | Score the paraffin section against its tissue IHC pattern and controls; do not require visible cilia or microtubules for a positive chromogenic call (HPA tissue IHC; general IHC practice). |
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). Use skeletal muscle myocytes as the tissue negative (HPA: Not detected in myocytes); on the adipose slide, cells without adipocyte staining should show only background, but are not independently validated negatives (HPA: High in adipocytes).
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 (caption: rabbit anti-AAMP PB9123; standard IHC practice). Add an AAMP knockout specimen or validated antigen-peptide block as a target-specific control, and quench endogenous peroxidase before HRP/DAB detection in adipose tissue (standard IHC practice; caption: HRP/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9123 paraffin-section caption does not state the fixative (caption: fixative not stated). That example uses heat-mediated EDTA retrieval at pH 8.0, but does not establish whether retrieval is required or frozen sections are easier (caption: EDTA pH 8.0). IF/ICC is not established as easier by the supplied evidence; HPA reports mainly microtubule localization, while lipid spaces in adipose sections can complicate interpretation (HPA: Microtubules approved; standard histology 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, compartment, cell type, and assay controls before comparing signal intensity.
How should I optimize retrieval when AAMP staining is weak?
Begin with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections stained with PB9123 (datasheet PB9123). The selected image used this retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C, so reproduce those conditions before changing retrieval (PB9123 tissue-IHC caption). If signal remains weak, compare shorter and longer heating runs on adjacent sections while holding antibody concentration and DAB development constant (standard IHC practice). Score signal in smooth muscle cells alongside background in a no-primary control; excessive heating can damage tissue morphology and make apparent gains in staining difficult to interpret (HPA: High in smooth muscle cells; standard IHC practice).
Could fixation explain inconsistent AAMP staining between paraffin blocks?
The selected paraffin-section caption does not report its fixative, so AAMP-specific fixation sensitivity is unknown (PB9123 tissue-IHC caption: fixative not stated). Record each block’s fixative and fixation time, then compare sections processed with the same pH 8.0 EDTA retrieval, 2 μg/ml primary concentration, and DAB development (datasheet PB9123; PB9123 tissue-IHC caption; standard IHC practice). Include a shared reference section in each run and examine whether weak staining follows particular blocks or the entire staining batch (standard IHC practice). Do not attribute a block difference to AAMP’s topology or phosphorylation without a direct fixation comparison using this antibody (UniProt Q13685: no transmembrane segment and phosphoserine at residue 20; standard IHC practice).
Which staining compartments should count as plausible AAMP signal?
For chromogenic tissue IHC, begin by evaluating cytoplasmic staining, particularly in smooth muscle cells, against local background (HPA tissue IHC: general cytoplasmic expression, most abundant in smooth muscle). UniProt also lists cell-membrane and cytoplasmic localisation, although AAMP has no annotated transmembrane segment; membrane-adjacent DAB staining therefore needs cellular context (UniProt Q13685: subcellular location and topology). Review high-power fields for a reproducible cellular pattern across intact regions rather than scoring diffuse pigment or section edges (standard IHC practice). HPA’s microtubule and ciliary assignments come from subcellular imaging and should not be assumed resolvable in routine DAB sections (HPA subcellular: microtubules and primary cilium; standard IHC practice).
How should an unknown AAMP epitope affect IHC optimization?
Treat the precise epitope as unmapped in the supplied evidence, and avoid assigning PB9123 staining to a particular AAMP region (PB9123 tissue-IHC caption: no epitope reported). UniProt lists 0 annotated isoforms, no annotated domains, and phosphorylation at serine 20; none establishes antibody binding to that residue (UniProt Q13685: isoforms, domains, modified residues). If staining changes after retrieval, compare adjacent sections under the same detection conditions before proposing epitope masking (datasheet PB9123: EDTA pH 8.0; standard IHC practice). Use an independently validated antibody with a documented, distinct epitope if regional specificity matters, and interpret agreement at the cell level rather than from DAB intensity alone (standard IHC practice).
How can IF help check an ambiguous AAMP IHC pattern?
Use IF/ICC as a separate validation experiment and multiplex AAMP with a smooth muscle marker when checking the cell population enriched in tissue IHC (HPA tissue IHC: most abundant in smooth muscle; standard IF practice). Select fluorophores after examining an unstained specimen, favoring channels with low tissue autofluorescence and minimal spectral overlap (standard IF practice). Because UniProt lists cytoplasmic and cell-membrane localisation but no transmembrane segment, the antibody’s epitope side is unresolved; compare permeabilized and nonpermeabilized preparations before interpreting surface signal (UniProt Q13685: localisation and topology; standard IF practice). HPA reports microtubule, cytosol, and plasma-membrane locations in subcellular imaging, which can guide IF pattern review without defining the chromogenic IHC endpoint (HPA subcellular: main and additional locations).
What controls help identify nonspecific brown staining in AAMP IHC?
Run a no-primary section through the same secondary reagent and DAB development to reveal detection-system or tissue background (standard IHC practice). The selected PB9123 image used 10% goat serum blocking, a peroxidase-conjugated secondary, and DAB; reproduce the block when comparing that workflow (PB9123 tissue-IHC caption). Add an endogenous peroxidase block and inspect pigment-rich, necrotic, and section-edge areas before calling brown deposits AAMP-positive (standard IHC practice). If background persists, shorten DAB development or titrate primary antibody around the documented 2 μg/ml, then compare specific cellular staining and background on matched sections (PB9123 tissue-IHC caption; standard IHC practice).
How should AAMP staining be scored across heterogeneous tissue? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, using cytoplasmic signal in identifiable cells as the primary tissue-IHC readout (HPA tissue IHC: general cytoplasmic expression; standard IHC practice). For each region, report the percentage of positive cells and an intensity-weighted H-score from 0–300, with thresholds fixed before reviewing groups (standard IHC practice). If the question concerns vessels or infiltrating cells, report positive-cell density per mm² of viable tissue and normalize to the relevant cell or vessel count (UniProt Q13685: expression in blood vessels; standard IHC practice). Exclude necrosis and section edges consistently, and keep retrieval, primary concentration, imaging, and DAB development comparable across slides (standard IHC practice).
How can I distinguish genuine AAMP staining from an artefact?
A credible result shows repeatable cellular staining in intact regions, with cytoplasmic signal and enrichment in smooth muscle providing tissue-IHC reference patterns (HPA tissue IHC: profile and High in smooth muscle cells; standard IHC practice). Interpret endothelial staining in anatomical context because AAMP is reported in blood vessels and strongly in endothelial cells (UniProt Q13685: tissue specificity). Be cautious with isolated nuclear deposits, necrotic areas, and strong section-edge staining when surrounding viable cells lack a consistent pattern (UniProt Q13685: cell membrane and cytoplasm; standard IHC practice). Check a no-primary control for endogenous peroxidase or detection background, and retain HPA’s Approved rating with its stated medium staining–RNA consistency as a limit on interpretation (standard IHC practice; HPA tissue IHC: reliability).
Boster reagents
Best AAMP / Angio-associated migratory cell protein IHC Antibodies
PB9123 has a human paraffin-section IHC figure (PB9123 image caption); M07409 lists IF/ICC for human, mouse and rat (M07409 applications and reactivity), with no IF figure supplied (M07409 image alts).
Real IHC data
Anti-AAMP Antibody ®
Cat # PB9123
The rendered PB9123 card shows AAMP staining in a paraffin-embedded human testis cancer section (PB9123 image caption). Its IHC dilution is 2–5 μg/ml for human samples (PB9123 dilution_raw); the caption does not report the fixative (PB9123 image caption).
Which to pick: Choose PB9123 for human paraffin-section IHC because its own figure documents that preparation and chromogenic detection (PB9123 image caption). For IF/ICC, M07409 lists those applications and a 1:50–1:200 dilution, but has no IF figure in the payload (M07409 applications, if_dilution and if_image_alts). Both list human, mouse and rat reactivity, while the supplied PB9123 IHC dilution specifies human and its IHC figure shows human tissue only (PB9123 and M07409 reactivity; PB9123 dilution_raw and image caption).
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 localisation 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 fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Skeletal myocytes show no detected staining despite reported expression (HPA tissue IHC; UniProt); Isoform / epitope: No annotated isoforms or cleavage; epitope sidedness unknown (UniProt); Regulation: Abundance varies by cell type (HPA tissue IHC). 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.