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AIP / AH receptor-interacting protein · IHC design guide
Design Immunohistochemistry for AIP
Plan AIP paraffin-section IHC around the cytoplasmic staining seen in most tissues (HPA tissue IHC). Use the catalog antibody at 0.5–1 μg/mL (datasheet M02759), and account for the unstained oral mucosa squamous cells when choosing controls (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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissue cell types (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 paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Oral mucosa squamous cells show no staining (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | No listed isoforms; single cytoplasmic chain, no ectodomain (UniProt) |
Section 1
Recommended AIP IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: M02759) is accompanied by four published AIP IHC protocols (PMC6432872; PMC6454377; PMC2862671; PMC9529452).
| 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 pH 8.0 retrieval for the catalog antibody (datasheet: M02759). Citrate pH 6.0 was used in one published protocol (PMC2862671: Methods).
Section 2
What Is the Expected AIP Staining Pattern?
AIP should stain mainly in the cytoplasm of many cell types (UniProt O00170: cytoplasm; HPA: cytoplasmic expression in most tissues). High staining is reported in adipocytes, bronchial respiratory epithelium and cerebral cortical neurons, among other cells (HPA: High). The HPA tissue IHC profile is Approved, with medium consistency between staining and RNA expression (HPA: reliability description). AIP has no transmembrane segment (UniProt O00170: topology).
| Clear cytoplasmic stain in bronchial respiratory epithelial cells or cerebral cortical neurons. | This fits the reported compartment and high staining in those cells (HPA: cytoplasmic profile; High in both cell types). Judge intensity against matched controls, since HPA levels describe observed tissue staining, not a required score for every specimen. |
| Predominantly nuclear or membrane outlined stain, with little cytoplasmic signal. | Recheck localisation before calling this AIP positive: HPA supports cytosol by ICC-IF and UniProt lists cytoplasm with no transmembrane segment (HPA: cytosol supported; UniProt O00170: subcellular location and topology). Review controls and detection. |
| Strong stain in oral mucosal squamous epithelium. | HPA reports AIP as Not detected in those cells (HPA: oral mucosa, squamous epithelial cells). Check antibody specificity and endogenous detection activity before interpreting unexpected staining; a single discrepant section does not prove cross-reactivity. |
| Uniform haze across cells and surrounding tissue, obscuring cell boundaries. | This is difficult to score as cytoplasmic AIP (HPA: cytoplasmic profile). Review reagent-only controls, blocking, washes and detection chemistry as general IHC checks; the supplied sources do not identify an AIP-specific cause of diffuse background. |
| No signal in a section expected to contain strongly staining cells. | First confirm that the expected cells are present: HPA reports High staining in adipocytes and adrenal glandular cells (HPA: tissue IHC). Then check the IHC-validated antibody, retrieval and detection controls before treating the absence as biological. |
💡Expected AIP appearanceA convincing positive shows predominantly cytoplasmic stain in the expected cells, including strong signal where HPA reports High staining; dominant nuclear, membrane outlined or uniform background stain calls for review (HPA: cytoplasmic profile and tissue IHC; UniProt O00170: topology).
| Tissue and cell context | AIP is widely expressed (UniProt O00170: tissue specificity), while HPA reports both High staining in selected cell types and Not detected in oral mucosal squamous cells (HPA: tissue IHC). Compare the named cells, not whole-section colour. |
| Liver as a comparator | UniProt says AIP is not expressed in liver, whereas HPA reports Low staining in cholangiocytes (UniProt O00170: tissue specificity; HPA: liver cholangiocytes, Low). Because these statements differ in scope and readout, avoid using liver as an unequivocal negative control. |
| IHC antibody evidence | Two listed mouse monoclonals have Approved IHC status, CAB033674 and CAB069477 (HPA: antibody validation). The tissue profile is Approved with medium RNA–staining consistency (HPA: reliability description); neither statement establishes that every observed signal is specific. |
| Protein location and processing | AIP is a 330-residue cytoplasmic protein without a transmembrane segment, signal peptide or reported cleavage into multiple chains (UniProt O00170: location, topology and processing). These annotations support an intracellular pattern, but do not locate the antibody epitope or predict retrieval performance. |
| IF/ICC Q&A | What should IF/ICC show? Predominantly cytosolic AIP (HPA: cytosol supported; UniProt O00170: cytoplasm). HPA lists HPA050217 as Supported for ICC and lists ICC-IF images in A-431, CACO-2, U2OS and NIH 3T3 (HPA: antibody validation and subcellular record). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected cytoplasmic stain is absent from a positive-control section. | The expected cells may be missing, or the IHC workflow may have failed; HPA reports High staining in adrenal glandular cells (HPA: tissue IHC). | Confirm cell identity and tissue preservation, then check retrieval, antibody application and chromogenic detection with appropriate run controls (general IHC practice). |
| Signal is mainly nuclear. | That distribution conflicts with the supported cytosolic location (HPA: subcellular location; UniProt O00170: cytoplasm). The image alone cannot identify the source of the mismatch. | Review compartment assignment against the counterstain and compare control sections; investigate antibody specificity and detection background before scoring (general IHC practice). |
| Oral mucosal squamous cells stain strongly. | This differs from HPA's Not detected observation for those cells (HPA: oral mucosa). Endogenous detection activity or nonspecific antibody staining is possible, but neither is established by this result. | Inspect reagent and antibody controls and repeat the cell-level assessment alongside an expected positive tissue (general IHC practice; HPA: tissue IHC). |
| A broad brown haze obscures cytoplasm. | Diffuse background prevents a reliable comparison with the expected cytoplasmic pattern (HPA: tissue IHC profile); its mechanism is unknown from these sources. | Check the no-primary control, blocking, wash steps and chromogen development, then score only cells with discernible localisation (general IHC practice). |
| A liver section gives weak focal staining. | HPA reports Low staining in cholangiocytes, while UniProt states that AIP is not expressed in liver (HPA: liver cholangiocytes; UniProt O00170: tissue specificity). | Identify the stained cell type and compare it with a clearer positive-control tissue; do not infer specificity or assay failure from liver alone. |
| The chosen antibody gives an unexpected tissue pattern. | HPA's Approved tissue profile has medium consistency with RNA data, and its listed IHC antibodies have Approved status (HPA: reliability description and antibody validation). Those ratings do not settle an individual discordant slide. | Compare cell localisation and controls with HPA's reported pattern; where available, use an independently validated antibody to investigate the discrepancy (general IHC practice). |
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: adipocytes should stain (HPA: High in adipocytes). Use oral mucosa as the negative tissue, assessing its squamous epithelial cells (HPA: Not detected in squamous epithelial cells); no cell type within the adipose slide is established as an internal AIP-negative control, so assess background in unstained areas without labeling other cells negative.
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 no-primary (secondary-only) and concentration-matched mouse IgG isotype controls (selected-SKU caption: mouse primary at 1 μg/ml), plus AIP-knockout material or peptide competition if suitable material is available. For chromogenic detection, block endogenous peroxidase and check for endogenous biotin if using the caption’s biotin-based detection method (selected-SKU caption: biotinylated secondary and streptavidin–biotin complex).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption leaves the fixative unreported. The caption demonstrates staining after heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU caption: paraffin section and EDTA retrieval). In adipose tissue, the thin adipocyte cytoplasmic rim can make cytoplasmic staining difficult to score (HPA: High in adipocytes; UniProt: Cytoplasm).
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
Use the catalog antibody’s paraffin section protocol as the starting point, then judge staining against AIP’s expected cytoplasmic distribution and tissue context.
Which retrieval conditions should I start with for AIP paraffin sections?
Start with heat mediated antigen retrieval in EDTA buffer at pH 8.0 (datasheet M02759). The catalog antibody’s paraffin section example used that retrieval before incubation at 1 µg/mL overnight at 4°C (datasheet M02759). If staining is weak, adjust heating duration or cooling gradually while keeping the buffer and detection steps constant (standard IHC practice). Compare each condition on adjacent sections with the same counterstain and exposure to chromogen, recording both cytoplasmic signal and tissue damage (standard IHC practice). AIP is reported in the cytoplasm, so assess recovery there rather than treating isolated nuclear colour as successful retrieval (UniProt O00170 subcellular location; HPA tissue IHC).
How should I troubleshoot weak staining when fixation history is uncertain?
The selected paraffin section caption does not report a fixative, so AIP specific sensitivity to fixation is unknown (datasheet M02759). Record the specimen’s fixative, fixation time and processing history before changing the antibody incubation or retrieval conditions (standard IHC practice). Compare sections with matched processing, starting with EDTA at pH 8.0 and the reported 1 µg/mL overnight incubation at 4°C (datasheet M02759). If signal differs, change one preparation variable at a time and inspect morphology alongside staining (standard IHC practice). Cytoplasmic distribution can guide interpretation, but it cannot establish how fixation affected this epitope (UniProt O00170 subcellular location; standard IHC practice).
What compartment should count as convincing AIP staining?
Score diffuse or granular cytoplasmic staining within intact cells as the expected pattern (UniProt O00170 subcellular location; HPA tissue IHC). The supported subcellular assignment is cytosol, and AIP has no transmembrane segment (HPA subcellular; UniProt O00170 topology). Check the pattern at the same microscope settings across adjacent sections, including the section incubated with 1 µg/mL primary antibody (datasheet M02759; standard IHC practice). Predominantly nuclear, crisp membrane or acellular staining needs independent validation before being called AIP (UniProt O00170 subcellular location; standard IHC practice). Use morphology and a counterstain to distinguish cytoplasm from overlapping cells or extracellular deposits (standard IHC practice).
Could an isoform or modified epitope explain inconsistent staining?
AIP has no annotated isoforms in the supplied record, so an isoform explanation is unsupported here (UniProt O00170 isoforms). Its FKBP type PPIase domain spans residues 31–121, and serine 43 is annotated as phosphorylated (UniProt O00170 domains; UniProt O00170 modified residues). The antibody epitope is not supplied, so neither the domain nor that modification establishes why a particular section stains weakly (datasheet M02759; standard IHC practice). Hold EDTA retrieval at pH 8.0 while comparing processing matched sections, then test one retrieval change at a time (datasheet M02759; standard IHC practice). Record whether any gain remains cytoplasmic and preserves tissue morphology (UniProt O00170 subcellular location; standard IHC practice).
How can IF help check the cell types and compartment seen by IHC?
Use IF as a separate check of the cytoplasmic pattern expected in chromogenic sections (HPA subcellular; HPA tissue IHC). Multiplex AIP with a marker for the cell type being assessed, such as an epithelial lineage marker when examining bronchus, where respiratory epithelial cells show high staining (HPA: High in bronchus respiratory epithelial cells; standard IF practice). Choose spectrally separated fluorophores and consider a far red channel if tissue autofluorescence obscures shorter wavelengths (standard IF practice). AIP is cytosolic and lacks a transmembrane segment, so use mild permeabilisation after fixation to reach intracellular epitopes, while checking that cell morphology remains intact (HPA subcellular; UniProt O00170 topology; standard IF practice).
How do I reduce diffuse brown staining without losing cytoplasmic AIP?
First compare a section processed without primary antibody against one stained at 1 µg/mL overnight at 4°C (datasheet M02759; standard IHC practice). The reported method blocked sections with 10% goat serum and used a biotinylated secondary, streptavidin biotin complex and DAB (datasheet M02759). Include a peroxidase block and inspect controls for endogenous enzyme activity or biotin related signal before attributing diffuse brown colour to AIP (standard chromogenic IHC practice). Optimise washing and chromogen development while retaining the same retrieval condition, EDTA at pH 8.0 (datasheet M02759; standard IHC practice). Count signal as credible only when cell associated cytoplasmic staining exceeds the control background (HPA tissue IHC; standard IHC practice).
What scoring method makes AIP IHC comparable across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring boundary before reviewing outcomes (HPA tissue IHC; standard IHC practice). For each section, report the percentage of positive cells and an H score based on the proportions at intensity grades 0–3; use positive cells per mm² when cell density is the question (standard IHC practice). Normalise counts to the number of evaluable cells or the measured viable tissue area, and exclude necrosis and torn edges consistently (standard IHC practice). Keep EDTA retrieval at pH 8.0, antibody concentration at 1 µg/mL, and chromogen development consistent between compared sections (datasheet M02759; standard IHC practice).
How can I separate true AIP positivity from artefact?
Look for cell associated cytoplasmic staining that follows intact morphology and exceeds the signal in a no primary control (UniProt O00170 subcellular location; standard IHC practice). HPA reports cytoplasmic expression in most tissues, with high staining in adipocytes and several glandular or neuronal populations, but no detection in oral mucosal squamous epithelial cells (HPA tissue IHC). Treat isolated nuclear or membrane colour, staining concentrated at cut edges or necrosis, and diffuse deposits as suspect until controls resolve them (HPA subcellular; standard IHC practice). DAB detection can also reflect endogenous peroxidase, so assess the peroxidase blocked control before interpreting brown colour (datasheet M02759; standard chromogenic IHC practice).
Boster reagents
Best AIP / AH receptor-interacting protein IHC Antibodies
Validated anti-AIP antibodies have IHC images from paraffin sections of human intestinal cancer, human tonsil and rat spleen (M02759 IHC captions), plus IF images from A431 and U20S cells (M02759 and PB9042 IF captions).
Real IHC data
Anti-ARA9/AIP Antibody ® (monoclonal, 10G8)
Cat # M02759
Real IF data
Anti-ARA9/AIP Antibody ®
Cat # PB9042
M02759 is listed for IHC and IF, with IHC images from human intestinal cancer, human tonsil and rat spleen paraffin sections and an IF image from A431 cells (M02759 applications and image captions). PB9042 is listed for IF and ICC, with an IF image from U20S cells; IHC is not listed for it (PB9042 applications and IF caption).
Which to pick: Choose M02759 for tissue IHC: its mouse monoclonal clone 10G8 is listed for IHC, and its IHC captions document heat retrieval in pH 8.0 EDTA on paraffin sections; the fixative is unreported (M02759 catalog entry and IHC captions). For IF/ICC, both SKUs are listed, with A431 IF data for M02759 and U20S IF data for rabbit polyclonal PB9042 (M02759 and PB9042 catalog entries and IF captions). For cross-species work, M02759 lists human, monkey, mouse and rat reactivity, while PB9042 lists human only (catalog reactivity).
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 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 paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Oral mucosa squamous cells show no staining (HPA tissue IHC); Isoform / epitope: No listed isoforms; single cytoplasmic chain, no ectodomain (UniProt); Regulation: No expression regulator specified (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.