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AADAT / Kynurenine/alpha-aminoadipate aminotransferase, mitochondrial · IHC design guide
Design Immunohistochemistry for AADAT
Plan chromogenic IHC in paraffin sections using hepatocytes and renal proximal tubules as positive tissue references (HPA tissue IHC). The guide pairs their cytoplasmic staining profile (HPA tissue IHC) with AADAT’s mitochondrial annotation (UniProt) and the catalog antibody’s 1:50–1:200 IHC dilution range (datasheet A09111).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AADAT 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 tissue staining (HPA tissue IHC); mitochondrial protein (UniProt) | |
| Staining pattern | Hepatocyte and proximal tubule cytoplasm; low GI staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A09111) | |
| Positive control | Kidney+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Only medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Higher expression in liver (UniProt) | |
| Isoform / epitope | 2 isoforms; mature chain 30–425; epitope coverage unknown (UniProt) |
Section 1
Recommended AADAT IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A09111) is paired with one published AADAT staining protocol for breast cancer tissue (PMC12874059).
| Sample | Paraffin-embedded human liver tissue; fixative not specified (datasheet A09111) |
| Fixation | Image fixative and duration unreported (datasheet A09111); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A09111); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-AADAT, 1:50-1:200 (datasheet A09111) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AADAT-positive staining in proximal tubules (cell body) of kidney (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in liver, renal tubules and low expression in the gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6.0 retrieval for the catalog antibody (datasheet A09111); the published protocol specifies pressure cooking for 10 minutes without a buffer (PMC12874059).
Section 2
What Is the Expected AADAT Staining Pattern?
For paraffin section IHC, expect cytoplasmic AADAT staining in hepatocytes and renal proximal tubule cells, with low staining in gastrointestinal glandular cells (HPA tissue IHC). Liver and kidney are medium level examples, and HPA rates the tissue IHC evidence Enhanced, with medium consistency between antibody staining and RNA data (HPA tissue IHC). UniProt places AADAT in mitochondria and annotates no transmembrane segment (UniProt Q8N5Z0).
| Hepatocytes and renal proximal tubule cell bodies show cytoplasmic chromogen, while adjacent structures remain distinguishable. | This matches the reported medium staining in both cell populations (HPA tissue IHC). A cytoplasmic result is compatible with mitochondrial localization (UniProt Q8N5Z0); ordinary chromogenic IHC need not resolve individual mitochondria (general IHC practice). |
| The dominant IHC signal is nuclear, with little cytoplasmic staining in hepatocytes or proximal tubules. | That distribution does not match the reported tissue IHC pattern (HPA tissue IHC) or mitochondrial annotation (UniProt Q8N5Z0). Treat it as a possible staining or interpretation artefact and assess controls before calling the cells AADAT positive (general IHC practice). |
| Strong staining appears mainly in cells expected to be negative, such as adipocytes or adrenal glandular cells. | Those cell types were not detected in the supplied tissue IHC record (HPA tissue IHC). Cross-reactivity or endogenous chromogenic detection activity are possibilities; neither is established by the image alone (general IHC practice). |
| Chromogen is widespread across cells, tissue spaces and section edges, obscuring cell boundaries. | Diffuse background prevents a reliable cell-specific call (general IHC practice). Compare background controls and the reported hepatocyte and proximal tubule pattern before assigning signal to AADAT (HPA tissue IHC). |
| A liver or kidney section has no discernible signal in hepatocytes or proximal tubules. | These are medium staining examples, so a blank result warrants a workflow check (HPA tissue IHC). It does not by itself prove absent AADAT: inspect tissue integrity, antibody and detection controls, and the run conditions (general IHC practice). |
💡Expected AADAT appearanceCall a positive IHC result when medium cytoplasmic staining identifies hepatocytes or renal proximal tubule cells (HPA tissue IHC); isolated nuclear signal or indiscriminate staining of expected negative cells is suspect (HPA tissue IHC; UniProt Q8N5Z0).
| Tissue and cell selection | Use liver hepatocytes or kidney proximal tubules as medium staining references; gastrointestinal glandular cells are low level examples (HPA tissue IHC). A weak gastrointestinal result is therefore less decisive than a blank liver or kidney result. |
| Compartment interpretation | UniProt annotates mitochondrial localization and no transmembrane segment (UniProt Q8N5Z0), while tissue IHC reports cytoplasmic staining (HPA tissue IHC). Assess chromogenic sections at the cellular level; do not require visible mitochondrial puncta (general IHC practice). |
| IF/ICC Q: Should its localization define this IHC pattern? | No. HPA ICC-IF reports approved plasma membrane localization and additional vesicles (HPA subcellular ICC-IF), which differs from the mitochondrial annotation (UniProt Q8N5Z0) and tissue IHC pattern (HPA tissue IHC). Interpret each assay with its own controls. |
| Strength of tissue evidence | HPA labels tissue IHC Enhanced but describes only medium consistency with RNA expression (HPA tissue IHC). Its antibody HPA037502 has Enhanced IHC and Approved ICC status (HPA antibodies); these ratings support interpretation without proving every signal specific. |
| Processing and isoforms | UniProt lists a mature chain spanning residues 30–425 and two isoforms (UniProt Q8N5Z0). The payload does not map an antibody epitope to either feature, so neither can predict a staining difference or justify a retrieval setting. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in the liver hepatocytes or kidney proximal tubules. | The observed result conflicts with medium staining in these cells (HPA tissue IHC); a failed staining run remains possible (general IHC practice). | Check section quality and run controls, then verify the antibody, detection reagents and applied dilution against their documented instructions (general IHC practice). |
| Nuclear staining dominates an otherwise weak section. | A nuclear pattern conflicts with cytoplasmic tissue IHC (HPA tissue IHC) and mitochondrial localization (UniProt Q8N5Z0). | Review the negative control and morphology; repeat with validated IHC conditions before scoring the nuclear signal as AADAT (general IHC practice). |
| Adipocytes or adrenal glandular cells stain as strongly as the intended positive cells. | Both cell types were reported as not detected (HPA tissue IHC); off-target binding or endogenous detection activity may contribute (general IHC practice). | Compare appropriate negative and detection controls, then judge whether staining remains cell specific in liver or kidney (HPA tissue IHC; general IHC practice). |
| Diffuse chromogen obscures the distinction between positive cells and background. | The source cannot identify a reagent-specific cause; nonspecific antibody binding or detection background are general IHC possibilities (general IHC practice). | Inspect no-primary and detection controls, then review blocking, washes and antibody concentration against the chosen validated workflow (general IHC practice). |
| Only a faint signal appears in gastrointestinal glandular cells. | Low expression in the gastrointestinal tract is reported for tissue IHC (HPA tissue IHC). | Use liver or kidney as a stronger reference before judging assay performance; keep faint gastrointestinal staining separate from background by checking controls (HPA tissue IHC; general IHC practice). |
| Membrane or vesicle signal seems to conflict with cytoplasmic IHC. | HPA ICC-IF reports plasma membrane and vesicle localization (HPA subcellular ICC-IF), whereas tissue IHC reports cytoplasm (HPA tissue IHC). | Record the assay and cell context, inspect controls, and avoid transferring an ICC-IF compartment call directly to a paraffin section IHC score (general IHC practice). |
Experiment evidence
IHC & IF images showing AADAT staining
A09111 · IHCImmunohistochemistry of paraffin-embedded human liver using AADAT Rabbit pAb at dilution of 1:200 .Perform high pressure antigen retrieval w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109576-AADAT/tissue/cerebral+cortex#imid_19774857
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109576-AADAT/tissue/colon#imid_19791119
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109576-AADAT/tissue/liver#imid_19791136
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109576-AADAT/tissue/kidney#imid_19791129
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037502; green = AADAT, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037502; green = AADAT, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AADAT IHC & IF
🧪Run liver first: hepatocytes must show staining (HPA: Medium in hepatocytes; selected tissue-IHC caption: human liver). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the liver slide, morphologically distinct nonhepatocyte cells should show no signal above the no-primary background, but are background comparators rather than validated antigen-negative cells (standard IHC practice).
Positive control tissue: Kidney (Proximal tubules (cell body), HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AADAT in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control for the catalog rabbit polyclonal antibody (selected tissue-IHC caption: Rabbit pAb; standard IHC practice); use AADAT knockout tissue, if available, as a biological negative (standard IHC practice). Quench endogenous peroxidase in liver before chromogenic detection, and block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected SKU A09111 tissue-IHC caption does not report a fixative. The paraffin-section example uses high-pressure retrieval in 10 mM citrate, pH 6.0, and antibody dilution 1:200 (selected tissue-IHC caption); retrieval dependence beyond that example is unreported. The evidence does not establish that frozen sections or IF are easier; liver endogenous peroxidase can cause chromogenic background (standard IHC practice), and reported IF localization differs between plasma membrane/vesicles and mitochondrion (HPA subcellular; UniProt Q8N5Z0).
HPA tissue IHC evidence for AADAT
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Kidney | Proximal tubules (cell body) | Medium | Protein (IHC) | HPA → |
| Liver | Hepatocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AADAT IHC Tips
Use liver and kidney staining patterns, compartment checks, and matched controls to troubleshoot chromogenic AADAT staining in paraffin sections (HPA tissue IHC; UniProt Q8N5Z0).
How should I adjust retrieval when AADAT staining is weak or uneven?
Start with heat-mediated retrieval in 10 mM citrate buffer, pH 6.0; the A09111 liver image used high-pressure retrieval before staining (datasheet A09111). Run a known positive liver section beside the test section, and compare hepatocyte staining at the catalog antibody’s illustrated 1:200 dilution (datasheet A09111; HPA tissue IHC). If staining remains weak, vary heating duration or cooling conditions one at a time while keeping tissue, detection, and imaging comparable (standard IHC practice). Check whether staining improves in hepatocyte cell bodies without creating diffuse edge staining or signal in the no-primary control (HPA tissue IHC; standard IHC practice).
Could fixation explain loss of AADAT staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the A09111 tissue caption identifies paraffin-embedded human liver but does not state its fixative (datasheet A09111). Record the specimen’s actual fixation method and duration, then compare sections with similar processing using the same 10 mM citrate, pH 6.0 retrieval (standard IHC practice; datasheet A09111). Include a liver control in each run and assess staining in hepatocyte cell bodies before changing antibody concentration (HPA tissue IHC; standard IHC practice). Differences between blocks can prompt a controlled processing comparison, but the available evidence cannot assign an AADAT-specific effect to fixation (datasheet A09111; standard IHC practice).
Why does my stain look membranous when AADAT is annotated as mitochondrial?
UniProt places AADAT in mitochondria and reports no transmembrane segment, whereas HPA tissue IHC describes cytoplasmic staining in liver and renal tubules (UniProt Q8N5Z0; HPA tissue IHC). HPA cell imaging instead lists plasma membrane as the main location and vesicles as an additional location, creating a compartment discrepancy to investigate (HPA subcellular). At matched exposure, compare hepatocyte or proximal-tubule cell-body signal with section edges, luminal surfaces, and no-primary staining (HPA tissue IHC; standard IHC practice). Treat a crisp membrane-only chromogenic pattern as provisional until it reproduces across sections and agrees with an independent localization control (UniProt Q8N5Z0; HPA subcellular; standard IHC practice).
Can epitope position or isoforms explain inconsistent AADAT staining?
AADAT has 2 annotated isoforms, while the supplied A09111 caption does not identify the antibody epitope or establish isoform coverage (UniProt Q8N5Z0; datasheet A09111). The annotated mature chain spans residues 30–425; lysine modifications include acetylation and alternative modifications at residue 263 (UniProt Q8N5Z0). Without an epitope map, neither processing nor those modifications can be assigned as the cause of a staining difference (UniProt Q8N5Z0; datasheet A09111). Compare matched sections under identical retrieval and detection, then use independently characterized epitope coverage if isoform recognition matters to the conclusion (standard IHC practice; UniProt Q8N5Z0).
How can I investigate AADAT localization by immunofluorescence alongside tissue IHC?
Treat IF as a separate assay: the A09111 evidence describes chromogenic staining of paraffin-embedded liver, with fixation unreported (datasheet A09111). Multiplex AADAT with an independently validated hepatocyte or proximal-tubule cell marker to identify the cells underlying tissue-IHC signal (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores and measure unstained-tissue autofluorescence, especially before interpreting weak cytoplasmic or vesicular signal (standard IF practice; HPA subcellular). Because AADAT is annotated as mitochondrial and has no transmembrane segment, choose permeabilisation that gives antibody access to intracellular epitopes; confirm the actual epitope’s accessibility experimentally (UniProt Q8N5Z0; standard IF practice).
What should I check when chromogenic AADAT staining appears widespread?
Compare test sections with a no-primary control and a liver reference, processed with identical retrieval and chromogen development (standard IHC practice; HPA tissue IHC). Block endogenous peroxidase before enzyme-based detection and check whether pigment, tissue edges, or necrotic areas retain colour without primary antibody (standard IHC practice). AADAT staining is reported in hepatocytes and proximal-tubule cell bodies, while several other sampled cell populations were not detected by HPA tissue IHC (HPA tissue IHC). If background persists, adjust antibody concentration and wash conditions one variable at a time; the A09111 liver image used 1:200 (standard IHC practice; datasheet A09111).
How should I score AADAT staining across liver or kidney sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring: HPA reports medium staining in hepatocytes and kidney proximal-tubule cell bodies (HPA tissue IHC). Within annotated regions, record the percentage of positive target cells and an H-score from intensity categories, using the same threshold and development conditions across slides (standard IHC practice). Normalize positive counts to the number of eligible target cells, or report positive-cell density per mm² of evaluable tissue when cell counts are impractical (standard IHC practice). Exclude necrosis, folds, and edge artefacts consistently, and report each specimen’s sampled regions rather than treating a field as an independent specimen (standard IHC practice).
When should I call an unexpected AADAT signal an artefact?
Give greatest weight to reproducible staining in hepatocytes or proximal-tubule cell bodies, the populations with medium HPA tissue-IHC signal (HPA tissue IHC). Flag isolated staining in an unexpected cell population, sharp section-edge signal, or colour concentrated in necrosis for control review (HPA tissue IHC; standard IHC practice). A membrane-only pattern needs further localization evidence because UniProt annotates mitochondria while HPA cell imaging reports plasma membrane and vesicles (UniProt Q8N5Z0; HPA subcellular). Compare no-primary and peroxidase-block controls to distinguish antibody-dependent staining from endogenous enzyme activity, then repeat any disputed pattern on an independently processed section (standard IHC practice).
Boster reagents
Best AADAT / Kynurenine/alpha-aminoadipate aminotransferase, mitochondrial IHC Antibodies
A09111 has a human liver paraffin-section IHC image (IHC image caption); IF/ICC is listed without an IF image (catalog applications; IF image alts). Reactivity lists human, mouse and rat (catalog reactivity).
Real IHC data
Anti-AADAT Antibody
Cat # A09111
A09111 is shown staining paraffin-embedded human liver at 1:200 after high-pressure retrieval in 10 mM citrate, pH 6.0 (IHC image caption). IHC and IF/ICC are listed applications, with human, mouse and rat reactivity; no IF image is supplied (catalog applications; catalog reactivity; IF image alts).
Which to pick: Choose A09111 for tissue IHC when a paraffin-section example is useful: its rabbit polyclonal antibody is shown on human liver (IHC image caption). A09111 is also the listed IF/ICC and cross-species option because its applications include IF and ICC and its reactivity lists human, mouse and rat; those uses have no accompanying IF image (catalog applications; catalog reactivity; IF image alts). The IHC caption specifies paraffin embedding and retrieval, but does not report the fixative (IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8N5Z0 (AADAT_HUMAN, Kynurenine/alpha-aminoadipate aminotransferase, mitochondrial).
- Human Protein Atlas. AADAT tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. AADAT subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to vesicles..
- Human Protein Atlas. AADAT antibody validation summary (1 antibodies).
- AADAT-Driven Metabolic Control of Malate and CoQ(10) Shapes Immune Evasion in Triple-Negative Breast Cancer. bioRxiv : the preprint server for biology 2026 — PMC12874059.
- PubMed PMID:12126930 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AADAT (Kynurenine/alpha-aminoadipate aminotransferase, mitochondrial). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); mitochondrial protein (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A09111). Validated IHC antibody: A09111 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Proximal tubules (cell body)); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver tissue; fixative not specified (datasheet A09111); 2. Fixation: Image fixative and duration unreported (datasheet A09111); 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: Citrate pH 6.0 (datasheet A09111); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-AADAT, 1:50-1:200 (datasheet A09111); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Only medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; mature chain 30–425 ; epitope coverage unknown (UniProt); Regulation: Higher expression in liver (UniProt). Evidence: UniProt Q8N5Z0, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A09111, PMC12874059. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AADAT/immunohistochemistry-ihc-guide.