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AIP / AH receptor-interacting protein · IHC design guide
Design Immunohistochemistry for AIP
Plan paraffin-section AIP IHC around the cytoplasmic pattern reported in most tissues (HPA tissue IHC). Compare high adipocyte staining with oral mucosal squamous epithelium, where AIP is not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AIP 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 staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Most tissues show cytoplasmic cell staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02759) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Oral mucosa |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Low liver cholangiocyte staining contrasts with UniProt (HPA tissue IHC; UniProt) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 0 isoforms listed; one unprocessed chain (UniProt) |
Section 1
Recommended AIP IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet M02759) with three published AIP IHC protocols (PMC6432872; PMC6454377; PMC2862671).
| Sample | Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet M02759) |
| Fixation | Image fixative and duration unreported (datasheet M02759); 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 M02759); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02759) |
| Primary antibody | Mouse monoclonal (clone 10G8) anti-AIP, 0.5-1μg/ml (datasheet M02759) |
| Primary incubation | Overnight at 4 °C (datasheet M02759) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet M02759) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AIP-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet M02759). Citrate retrieval at pH 6.0 is a published alternative (PMC2862671).
Section 2
What Is the Expected AIP Staining Pattern?
AIP is a cytoplasmic protein with no transmembrane segment (UniProt O00170: subcellular location and topology). In paraffin sections, expect cytoplasmic staining across many tissues, with intensity judged in the relevant cell type: HPA reports high staining in adipocytes and several epithelial and neuronal populations (HPA tissue IHC). HPA rates its tissue pattern Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic stain in adrenal glandular cells or cerebellar Purkinje cells. | This fits the reported compartment and high staining in those cell types (UniProt O00170: cytoplasm; HPA tissue IHC: High). Compare cells within the section rather than treating every cell as an equally strong positive (HPA tissue IHC: cell-specific levels). |
| Predominantly nuclear, membranous, or extracellular stain with little cytoplasmic signal. | That distribution does not match the reported AIP location (UniProt O00170: cytoplasm; HPA subcellular: supported cytosol). Review morphology and staining controls before interpreting it as a new AIP location; AHR signaling alone does not establish nuclear AIP staining (UniProt O00170: function and subcellular location). |
| Strong stain in oral mucosa squamous cells, despite an otherwise plausible slide. | HPA reports AIP as not detected in those cells, while cervix squamous cells stain highly (HPA tissue IHC). Check cell identification, nonspecific binding, and endogenous detection activity (standard IHC practice); squamous identity alone cannot predict AIP positivity (HPA tissue IHC). |
| Uniform haze across tissue, stroma, and empty areas obscures cell boundaries. | A broad haze cannot establish the cytoplasmic, cell-resolved pattern reported for AIP (HPA tissue IHC; HPA subcellular). Assess background with an appropriate negative detection control, then review blocking, washes, and antibody concentration (standard IHC practice). |
| No stain in a section expected to contain high-expressing cells. | Absent signal in HPA-high cells, such as adrenal glandular cells, leaves the assay unverified for that run (HPA tissue IHC: High). Check that the cells are present and that the primary antibody, retrieval, and chromogenic detection steps worked (standard IHC practice). |
💡Expected AIP appearanceCall a result positive when stain is predominantly cytoplasmic in identifiable HPA-high cells, such as adrenal glandular cells or cerebellar Purkinje cells; isolated nuclear or diffuse tissue-wide color is suspect (UniProt O00170: cytoplasm; HPA tissue IHC: High; HPA subcellular: supported cytosol).
| Compartment and topology | AIP is cytoplasmic and lacks a transmembrane segment (UniProt O00170); HPA independently supports a cytosolic IF location (HPA subcellular). Score intracellular cytoplasmic color in intact cells (standard IHC practice). |
| Tissue and cell context | HPA reports high staining in adipocytes, adrenal glandular cells, and Purkinje cells, but none detected in oral mucosa squamous cells (HPA tissue IHC). Choose and interpret controls at the cell level (standard IHC practice). |
| Liver evidence | UniProt says AIP is not expressed in liver, while HPA reports low staining in cholangiocytes (UniProt O00170: tissue specificity; HPA tissue IHC: Low). Treat liver as an uncertain negative comparator, not a guaranteed blank section. |
| Antibody evidence | HPA lists two mouse monoclonals with Approved IHC status, CAB033674 and CAB069477; the listed rabbit polyclonal HPA050217 has Supported ICC status but no IHC status (HPA antibodies). These statuses do not establish identical IHC patterns across antibodies. |
| Retrieval and chromogenic detection | No AIP-specific retrieval condition or fixation sensitivity is supplied by UniProt or HPA. Retrieval adequacy must be judged with assay controls; endogenous enzyme activity can create misleading chromogenic color (standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No cytoplasmic signal in an expected positive section. | A failed staining step or absence of the relevant cells can mimic a negative result (standard IHC practice). | Confirm HPA-high cells are present, then review the primary antibody, retrieval, detection reagents, and run controls (HPA tissue IHC: High; standard IHC practice). |
| Only nuclei stain. | The observed compartment conflicts with cytoplasmic AIP and supported cytosolic IF staining (UniProt O00170; HPA subcellular). | Check counterstain and section morphology, then repeat with suitable negative controls before assigning AIP positivity (standard IHC practice). |
| Unexpected strong stain in oral mucosa squamous cells. | This conflicts with HPA's not-detected result; nonspecific binding or endogenous activity is possible (HPA tissue IHC; standard IHC practice). | Compare a primary-omission control and examine whether color follows cells or background (standard IHC practice). |
| Diffuse brown background masks cell boundaries. | Excess background can prevent reliable compartment scoring (standard IHC practice). | Review blocking, washes, chromogen development, and antibody concentration against the run's controls (standard IHC practice). |
| Liver shows faint cholangiocyte stain. | HPA records low cholangiocyte staining despite UniProt's liver nonexpression statement (HPA tissue IHC: Low; UniProt O00170: tissue specificity). | Record the stained cell type and intensity; use a clearer HPA-high tissue to judge assay performance (HPA tissue IHC; standard IHC practice). |
| IF/ICC question: where should AIP appear? | The IF location is a supported cytosolic signal, including HPA cell-line images (HPA subcellular). | Interpret cytosolic fluorescence against an appropriate background control (HPA subcellular; standard IF practice). Consult the separate IF/ICC guide for assay setup. |
Experiment evidence
IHC & IF images showing AIP staining
M02759 · IHCIHC analysis of ARA9/AIP using anti-ARA9/AIP antibody (M02759). ARA9/AIP was detected in paraffin-embedded section of human intestinal cance
M02759 · IHCIHC analysis of ARA9/AIP using anti-ARA9/AIP antibody (M02759). ARA9/AIP was detected in paraffin-embedded section of human tonsil tissue. H
M02759 · IFIF analysis of ARA9/AIP using anti-ARA9/AIP antibody (M02759). ARA9/AIP was detected in immunocytochemical section of A431 cells. Enzyme ant
PB9042 · IFIF analysis of ARA9 using anti-ARA9 antibody (PB9042). ARA9 was detected in immunocytochemical section of U20S cells. Enzyme antigen retriev
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000110711-AIP/tissue/cerebral+cortex#imid_18699472
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000110711-AIP/tissue/colon#imid_18699390
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000110711-AIP/tissue/liver#imid_18699399
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000110711-AIP/tissue/pancreas#imid_18699334
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA050217; green = AIP, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA050217; green = AIP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AIP IHC & IF
🧪Run adipose tissue first and score adipocytes for AIP staining (HPA: High in adipocytes). Run oral mucosa as the negative tissue (HPA: Not detected in squamous epithelial cells); on the adipose slide, treat unstained nonadipocyte cells as internal comparators only after confirming their identity and staining pattern, because the supplied adipose row reports adipocytes only (HPA positive row: adipocytes).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Oral mucosa (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AIP in A-431, CACO-2, U2OS, NIH 3T3, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched mouse IgG isotype control for the catalog antibody (hero caption: mouse anti-AIP; standard IHC practice); use confirmed AIP-knockout material as a biological negative (standard IHC practice). For chromogenic detection, quench endogenous peroxidase and check for endogenous biotin if using the caption’s biotin-based system (hero caption: biotinylated secondary and SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU caption does not state the fixative (M02759 caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the caption’s paraffin-section example, but retrieval dependence in adipose tissue is unreported (M02759 caption). The supplied evidence does not establish whether frozen sections or IF are easier; in adipose sections, lipid extraction can leave empty-looking adipocytes that require careful cell identification (standard histology practice).
HPA tissue IHC evidence for AIP
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 → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AIP IHC Tips
Troubleshoot AIP staining in paraffin sections by checking retrieval, cytoplasmic localisation, controls and cell-specific scoring before interpreting signal.
Which retrieval condition should I start with for AIP in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M02759). The selected tissue image used that condition before overnight incubation with 1 μg/ml antibody at 4°C (datasheet M02759). If staining is weak, compare retrieval duration on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Include a known positive tissue section in each comparison, since AIP staining is cytoplasmic in most tissues (HPA: tissue IHC profile). Record tissue morphology alongside signal intensity; excessive heating can damage sections and complicate interpretation (standard IHC practice).
Can fixation explain weak or uneven AIP staining?
Target-specific fixation sensitivity for AIP is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (datasheet M02759). Record the actual fixative and fixation interval for each specimen before comparing staining across cases (standard IHC practice). If sections differ in processing history, assess them with the same EDTA pH 8.0 retrieval and 1 μg/ml starting antibody concentration (datasheet M02759; standard IHC practice). Check morphology and an internal positive cell population before attributing weak signal to fixation (standard IHC practice). Do not infer fixation tolerance from AIP tissue distribution or protein features (UniProt O00170; HPA: tissue IHC profile).
What staining pattern is plausible for AIP, and when should nuclear signal concern me?
Expect predominantly cytoplasmic staining in paraffin sections (UniProt O00170: cytoplasm; HPA: cytoplasmic expression in most tissues). Independent cell imaging places AIP in the cytosol (HPA: supported cytosol location). Score cytoplasmic signal separately from nuclear or stromal colour, using the same counterstain and microscope settings across sections (standard IHC practice). If nuclear staining dominates, review the negative reagent control and compare a section processed with the documented EDTA pH 8.0 retrieval (datasheet M02759; standard IHC practice). AIP has no transmembrane segment, so a crisp membrane-only pattern needs independent confirmation (UniProt O00170: topology).
How should I investigate an unexpected AIP staining pattern without an epitope map?
The supplied record lists 0 isoforms and a 330-residue AIP chain (UniProt O00170: isoforms and processing). It identifies an FKBP-type PPIase domain at residues 31–121 and a phosphoserine at residue 43, but provides no antibody epitope (UniProt O00170; datasheet M02759: epitope not supplied). Do not assign staining differences to domain masking or phosphorylation without epitope and validation data (standard IHC interpretation). Compare serial sections under identical EDTA pH 8.0 retrieval and detection conditions, including a negative reagent control (datasheet M02759; standard IHC practice). If the pattern remains unexpected, seek independent AIP-specific validation before calling a distinct molecular form (standard IHC practice).
How can I check AIP localisation by IF alongside chromogenic IHC?
Use IF as a separate localisation check: AIP is cytoplasmic and has no transmembrane segment (UniProt O00170: subcellular location and topology). Pair the AIP channel with a validated marker for the expected cell type, such as adipocytes in adipose tissue, and assess channel overlap at the cell level (HPA: high AIP in adipocytes; standard IF practice). Permeabilise cells or sections to reach a cytosolic epitope, then optimise treatment because the antibody epitope is unspecified (HPA: supported cytosol location; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section for tissue autofluorescence (standard IF practice). The paraffin IHC caption supplies no IF fixation or dilution condition (datasheet M02759).
How do I reduce diffuse brown staining without losing AIP signal?
First compare stained sections with a negative reagent control to locate background from detection chemistry (standard IHC practice). The selected paraffin-section image used 10% goat serum blocking, 1 μg/ml primary antibody overnight at 4°C, and biotinylated secondary detection with DAB (datasheet M02759). Apply a peroxidase block before DAB development and consider a biotin-related control for this detection system (standard chromogenic IHC practice; datasheet M02759: biotinylated detection). Shorten chromogen development or titrate antibody if diffuse colour obscures cells, changing one variable at a time (standard IHC practice). Preserve interpretable cytoplasmic signal when judging improvement (HPA: tissue IHC profile).
How should I quantify AIP across sections with different cell compositions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, because AIP expression varies by tissue and cell type (HPA: tissue IHC profile). For cytoplasmic DAB staining, report the percentage of positive target cells and an H-score calculated as the sum of each intensity grade multiplied by its percentage, yielding 0–300 (HPA: cytoplasmic tissue expression; standard IHC scoring). Normalise counts to the number of evaluable target cells; for spatial comparisons, report positive cells per mm² of viable tissue (standard IHC practice). Keep retrieval, antibody concentration, development and scoring thresholds consistent across sections (standard IHC practice). Exclude folds, edges and necrotic areas from the scored region (standard IHC practice).
When is apparent AIP positivity more likely to be artefact?
A plausible positive result is cytoplasmic staining in identifiable cells; AIP is reported in the cytoplasm across most tissues (UniProt O00170: location; HPA: tissue IHC profile). Compare the stained cell type with the section's anatomy: HPA reports high signal in adipocytes and adrenal glandular cells, but no detected signal in oral mucosa squamous epithelial cells (HPA: tissue IHC examples). Treat isolated edge staining, necrotic deposits or colour reproduced in a negative reagent control as suspect (standard IHC practice). Check endogenous peroxidase contribution when DAB persists without primary antibody (standard chromogenic IHC practice). HPA rates tissue staining as approved with medium RNA consistency, so resolve discordant cases with independent validation (HPA: reliability description).
Boster reagents
Best AIP / AH receptor-interacting protein IHC Antibodies
Catalog anti-AIP antibodies have IHC data from human intestinal cancer, human tonsil, and rat spleen paraffin sections (M02759 image captions), plus IF data from A431 and U20S cells (M02759 and PB9042 image captions).
Real IHC data
Anti-ARA9/AIP Antibody ® (monoclonal, 10G8)
Cat # M02759
Real IF data
Anti-ARA9/AIP Antibody ®
Cat # PB9042
M02759 will render with IHC data from human intestinal cancer paraffin sections; its catalog also lists IHC and IF and reactivity with human, monkey, mouse, and rat (M02759 image caption; catalog applications and reactivity). PB9042 will render with IF data from U20S cells; its catalog lists IF/ICC and human reactivity, with no IHC application (PB9042 image caption; catalog applications and reactivity).
Which to pick: Choose M02759 for paraffin-section IHC: its monoclonal antibody has IHC data from human intestinal cancer, human tonsil, and rat spleen sections (M02759 catalog clone; IHC image captions). For IF/ICC, PB9042 has human-cell IF data, while M02759 also has A431-cell IF data (PB9042 and M02759 IF image captions). M02759 offers the broader listed species reactivity—human, monkey, mouse, and rat—but its IHC captions establish processing for paraffin sections only; the fixative is unreported (M02759 catalog reactivity; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00170 (AIP_HUMAN, AH receptor-interacting protein).
- Human Protein Atlas. AIP tissue IHC expression (reliability: Approved).
- Human Protein Atlas. AIP subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. AIP antibody validation summary (3 antibodies).
- AIP and MEN1 mutations and AIP immunohistochemistry in pituitary adenomas in a tertiary referral center. Endocrine connections 2019 — PMC6432872.
- AIP-mutated acromegaly resistant to first-generation somatostatin analogs: long-term control with pasireotide LAR in two patients. Endocrine connections 2019 — PMC6454377.
- Isolated familial somatotropinoma: 11q13-loh and gene/protein expression analysis suggests a possible involvement of aip also in non-pituitary tumorigenesis. Clinics (Sao Paulo, Brazil) 2010 — PMC2862671.
- Association of Pathology Markers with Somatostatin Analogue Responsiveness in Acromegaly. International journal of endocrinology 2022 — PMC9529452.
- PubMed PMID:8972861 — UniProt-cited evidence.
- PubMed PMID:9111057 — UniProt-cited evidence.
- PubMed PMID:16728643 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AIP (AH receptor-interacting protein). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M02759). Validated IHC antibody: M02759 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Oral mucosa. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet M02759); 2. Fixation: Image fixative and duration unreported (datasheet M02759); 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 M02759); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M02759); 8. Primary antibody: Mouse monoclonal (clone 10G8) anti-AIP, 0.5-1μg/ml (datasheet M02759); 9. Primary incubation: Overnight at 4 °C (datasheet M02759). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Low liver cholangiocyte staining contrasts with UniProt (HPA tissue IHC; UniProt); Isoform / epitope: 0 isoforms listed; one unprocessed chain (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt O00170, HPA tissue IHC · Approved, HPA ICC-IF · cytosol., Datasheet M02759, PMC6432872, PMC6454377. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AIP/immunohistochemistry-ihc-guide.