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AKR1A1 / Aldo-keto reductase family 1 member A1 · IHC design guide
Design Immunohistochemistry for AKR1A1
Plan AKR1A1 chromogenic IHC in paraffin sections using renal tubular cells as a high-staining reference (HPA tissue IHC). This guide covers fixation, retrieval, antibody titration and interpretation of the observed cytoplasmic and nuclear pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AKR1A1 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 | Renal tubules show cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic and nuclear staining in renal tubules (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Caudate+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | No isoforms annotated; processed chain starts at residue 2 (UniProt) |
Section 1
Recommended AKR1A1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is supplemented by four published AKR1A1 IHC protocols from mouse liver and kidney sections (PMC10837880; PMC5352327; PMC4834580; PMC6318002).
| Sample | Paraffin-embedded human tonsil carcinoma tissue; fixative not specified (datasheet A04855) |
| Fixation | Image fixative and duration unreported (datasheet A04855); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-AKR1A1, 1:50-1:200 (datasheet A04855) |
| 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 | AKR1A1-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and nuclear expression in renal tubules. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); the published liver protocols use longer retrieval (PMC5352327; PMC4834580).
Section 2
What Is the Expected AKR1A1 Staining Pattern?
AKR1A1 is mainly cytosolic, with an additional nucleoplasmic ICC-IF signal and an apical membrane annotation; it has no transmembrane segment (HPA: subcellular; UniProt P14550: localization and topology). In paraffin IHC, expect the strongest reported staining in renal tubular cells, with medium staining in hepatocytes and lung macrophages (HPA: tissue IHC). HPA rates the tissue profile Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Renal tubular cells show prominent cytoplasmic staining, with some nuclear staining. | This fits the reported high tubular signal and cytoplasmic and nuclear renal pattern (HPA: kidney IHC). Interpret the nuclear component alongside the cytosolic localization and additional nucleoplasmic ICC-IF signal (HPA: subcellular). |
| Hepatocytes or lung macrophages stain less strongly than renal tubules. | Medium staining is reported in those specific cell populations, versus high staining in kidney tubular cells (HPA: tissue IHC). Score the named cells; a tissue-level label does not imply that every cell should stain. |
| Extracellular deposits or a compartment alone dominate the slide. | A dominant extracellular pattern does not match the reported cellular localization (UniProt P14550: cytosol and apical membrane; HPA: subcellular). Exclusively nuclear staining is also questionable because HPA describes AKR1A1 as mainly cytosolic (HPA: subcellular). Reassess specificity before scoring either pattern as positive. |
| Squamous epithelium or other unexpected cells stain as strongly as renal tubules. | HPA reports AKR1A1 as undetected in cervical, esophageal and oral squamous epithelial cells (HPA: tissue IHC). Unexpected chromogen may reflect cross-reactivity or endogenous detection activity; staining alone cannot distinguish them (general IHC practice). |
| A uniform haze covers cells and surrounding tissue, obscuring cell boundaries. | This cannot establish the reported cell-specific pattern (HPA: kidney IHC). Diffuse background can arise from nonspecific antibody binding or the detection system (general IHC practice); compare controls before assigning an AKR1A1 score. |
💡Expected AKR1A1 appearanceCall a result positive when renal tubular cells show high, chiefly cytoplasmic staining, with a possible nuclear component (HPA: kidney IHC; HPA: subcellular); medium hepatocyte or lung macrophage staining is also reported (HPA: tissue IHC). Diffuse extracellular chromogen or uniformly strong staining of HPA-undetected cells is suspect (HPA: tissue IHC; general IHC practice).
| Cellular localization | Cytosol is the main supported ICC-IF location; nucleoplasm is an additional approved location (HPA: subcellular). UniProt also annotates apical cell membrane, despite no transmembrane segment (UniProt P14550: localization and topology). Do not require a membrane rim to accept an otherwise matching renal pattern. |
| Choice of tissue and cells | Kidney tubular cells provide the clearest reported high-signal reference (HPA: kidney IHC). Liver hepatocytes and lung macrophages are medium (HPA: tissue IHC). HPA's undetected calls apply to named cell types, so a negative call in one cell type does not make its entire tissue a negative control. |
| Evidence and antibody choice | HPA rates the tissue profile Approved but notes medium staining–RNA consistency (HPA: tissue IHC). Four listed antibodies have Approved IHC status (HPA: antibodies). Match the actual IHC-validated antibody and its controls to the experiment; those ratings do not validate every reagent or every stained cell. |
| Retrieval and chromogenic detection | Use the catalog antibody's IHC-P instructions to select retrieval and dilution, then check controls (general IHC practice). No AKR1A1-specific fixation or retrieval sensitivity is supplied here; neither HPA tissue levels nor UniProt topology establish one. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in kidney tubular cells. | The high-signal reference has failed (HPA: kidney IHC); ineffective retrieval, antibody application or detection are possible workflow causes (general IHC practice). | Check section integrity and a known-positive control; verify retrieval, primary antibody use and chromogen steps against the IHC-P instructions (general IHC practice). Do not infer that AKR1A1 is absent from the specimen from this slide alone. |
| Only nuclear staining appears in renal tubules. | Renal nuclear staining is reported, but HPA describes AKR1A1 as mainly cytosolic (HPA: kidney IHC; HPA: subcellular). A nuclear-only result therefore needs confirmation. | Compare the same run with a renal positive control and a primary-omission control; reassess compartment assignment and reagent specificity before scoring (general IHC practice). |
| Strong staining appears in HPA-undetected squamous cells. | This conflicts with reported undetected squamous epithelial cells in cervix, esophagus and oral mucosa (HPA: tissue IHC). Cross-reactivity or endogenous chromogenic activity is possible (general IHC practice). | Check a primary-omission control and appropriate detection controls; compare the unexpected cells with renal tubular staining before treating them as AKR1A1-positive (general IHC practice; HPA: kidney IHC). |
| Background obscures the tubular pattern. | Nonspecific binding or residual endogenous enzyme activity may contribute to chromogenic background (general IHC practice). The image cannot reliably resolve the HPA-reported renal cell pattern (HPA: kidney IHC). | Review blocking and detection controls, antibody dilution, washing and chromogen development under the IHC-P workflow; score only interpretable cells (general IHC practice). |
| A proposed negative tissue contains some positive cells. | HPA's negative entries identify particular cell types, while its profile includes staining across multiple tissues (HPA: tissue IHC). A whole section may therefore be an ambiguous negative comparator. | Assess the specified HPA-undetected cell population, such as esophageal squamous epithelium, rather than calling every cell in that section negative (HPA: tissue IHC). Include a renal tubular positive reference in the run (HPA: kidney IHC). |
| Can an IF/ICC image settle an ambiguous IHC pattern? | HPA ICC-IF supports mainly cytosolic and additional nucleoplasmic localization; it does not directly validate chromogenic paraffin staining in the examined section (HPA: subcellular; general IHC practice). | Use the ICC-IF localization as a compartment cross-check and judge the IHC result against tissue-specific staining and its own controls (HPA: subcellular; HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing AKR1A1 staining
A04855 · IHCImmunohistochemistry (IHC) analyzes of AKR1A1 pAb in paraffin-embedded human tonsil carcinoma tissue at 1:50,showing cytoplasmic and nuclear
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000117448-AKR1A1/tissue/cerebral+cortex#imid_4663216
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000117448-AKR1A1/tissue/cerebellum#imid_4663211
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000117448-AKR1A1/tissue/colon#imid_4663134
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000117448-AKR1A1/tissue/liver#imid_4663143
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA017919; green = AKR1A1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA017919; green = AKR1A1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AKR1A1 IHC & IF
🧪Run kidney first: tubular cells should stain strongly (HPA: Kidney, cells in tubules, High). Use cervix squamous epithelium as a negative tissue (HPA: Cervix, squamous epithelial cells, Not detected); on the kidney slide, treat cells outside the positive tubules as internal negatives only if they remain at background, without assuming a particular cell type is negative.
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AKR1A1 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and normal rabbit IgG matched to the catalog polyclonal primary as an isotype control (caption: pAb; Goat Anti-Rabbit IgG-biotin secondary). Use knockout-validated material or an immunizing-peptide block, if available, as a biological specificity control; check kidney sections for endogenous peroxidase and biotin signal when using the captioned avidin–peroxidase detection (caption: avidin-peroxidase).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the selected A04855 paraffin-section caption does not state a fixative, so it does not establish fixation (A04855 caption: paraffin-embedded tissue; fixative not stated). The supplied evidence does not establish that frozen sections or IF are easier, although ICC-IF images support predominantly cytosolic localization (HPA subcellular: Cytosol, supported). In kidney, assess endogenous peroxidase and biotin as potential detection artefacts when using the captioned avidin–peroxidase system (A04855 caption: avidin-peroxidase). The selected A04855 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A04855).
HPA tissue IHC evidence for AKR1A1
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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AKR1A1 IHC Tips
Troubleshoot AKR1A1 staining in paraffin sections using the catalog image, tissue expression evidence, and controls appropriate for chromogenic IHC.
What retrieval should I try first for weak AKR1A1 staining in paraffin sections?
Use citrate buffer at pH 6.0 for heat induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule). Let sections cool in buffer before washing, and process a kidney positive control alongside the test section (standard IHC practice; HPA: high staining in renal tubule cells). If staining remains weak, compare a carefully controlled alternative retrieval buffer on serial sections while keeping antibody dilution and detection constant (standard IHC practice). Check tissue preservation and the primary omission control before attributing a stronger signal to improved epitope exposure (standard IHC practice).
Could fixation explain inconsistent AKR1A1 staining across paraffin samples?
AKR1A1 specific sensitivity to fixation is unknown from the supplied evidence; the catalog image describes paraffin sections but does not state a fixative (caption A04855). Record each specimen's fixative, fixation duration, and processing history before comparing staining intensity across cases (standard IHC practice). Run sections with known processing histories through the same pH 6.0, 95–98 °C, 20 min retrieval workflow and include a kidney control (page retrieval rule; HPA: high staining in renal tubule cells). If results differ, test matched processing conditions before assigning the variation to AKR1A1 abundance (standard IHC practice).
How should I assess nuclear or apical staining alongside cytoplasmic AKR1A1?
Score cytoplasmic staining first because AKR1A1 is mainly cytosolic; nucleoplasmic signal is also reported (HPA subcellular: supported cytosol, approved nucleoplasm). Apical membrane staining is plausible from the protein annotation, although AKR1A1 has no transmembrane segment (UniProt P14550: subcellular location and topology). The catalog paraffin image reports cytoplasmic and nuclear staining in tonsil carcinoma at 1:50, without establishing that every nuclear signal is specific (caption A04855). Compare each compartment with kidney tubules and a primary omission control, then record whether staining follows cell boundaries or appears only at section edges (HPA: high in renal tubules; standard IHC practice).
How can epitope uncertainty affect interpretation of AKR1A1 staining?
No AKR1A1 isoforms are annotated, and the reported mature chain spans residues 2–325 (UniProt P14550: isoforms and processing). The supplied evidence does not map the catalog antibody's epitope, so staining differences cannot be assigned to a particular sequence region (catalog antibody evidence). AKR1A1 has reported modifications near the N terminus and at residue 127; their effect on this antibody's IHC staining is unknown (UniProt P14550: modified residues). If epitope identity matters, obtain the antibody's immunogen information and compare an independently mapped antibody on adjacent sections under matched retrieval conditions (standard IHC validation practice).
How can IF help resolve ambiguous AKR1A1 chromogenic staining?
Use IF on a separately validated preparation to compare AKR1A1 with a marker identifying the expected cell type, such as a renal tubule marker in kidney (HPA: high staining in renal tubule cells; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence in unstained and secondary only controls, favouring a spectrally distinct channel where background is lower (standard IF practice). Because AKR1A1 is mainly cytosolic and has no transmembrane segment, include permeabilisation when testing intracellular staining, with a matched condition to assess its effect (HPA subcellular; UniProt P14550 topology; standard IF practice). Keep IF localisation conclusions separate from the paraffin IHC result (standard assay validation practice).
What should I check when AKR1A1 staining is diffuse or unexpectedly widespread?
Start with a primary omission control and assess whether the signal persists after the primary antibody is removed; the catalog image includes a PBS instead of primary control (caption A04855). Quench endogenous peroxidase before chromogenic detection, then examine background from the secondary reagent and the DAB reaction (standard IHC practice). Titrate the catalog antibody around the reported 1:50 image condition on serial sections, keeping retrieval and exposure to DAB consistent (caption A04855; standard IHC practice). Judge residual staining against expected kidney tubule enrichment and compartment patterns, while allowing for broader AKR1A1 expression (HPA tissue IHC; HPA subcellular).
How should I quantify AKR1A1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score cytoplasmic and nuclear staining separately before comparing specimens (HPA subcellular: cytosol and nucleoplasm; standard IHC scoring practice). An H-score sums the percentage of cells at each intensity multiplied by its 0–3 intensity grade, yielding 0–300; also report the percentage of positive cells (standard IHC scoring practice). For spatial analyses, report positive cell density per mm² of viable tissue or a defined compartment, and normalise against the number of eligible cells or measured viable area (standard IHC quantification practice). Keep retrieval, detection, counterstain, imaging, and scoring thresholds consistent, using kidney tubules as a staining reference (page retrieval rule; HPA: high in renal tubules; standard IHC practice).
How can I distinguish genuine AKR1A1 staining from artefact?
Give greatest weight to reproducible cytoplasmic staining in the expected cells, especially renal tubules; nucleoplasmic and apical patterns also have supporting annotations (HPA tissue IHC; HPA subcellular; UniProt P14550 subcellular location). Flag staining confined to section edges, folds, or necrotic regions for review against adjacent intact tissue (standard IHC practice). Compare unexpected cell types with HPA patterns, including high renal tubule staining and undetected cardiomyocyte staining, without treating those patterns as absolute exclusion rules (HPA tissue IHC). Persistent colour in a primary omission control points to background from detection chemistry or endogenous activity rather than demonstrated AKR1A1 binding (caption A04855; standard IHC practice).
Boster reagents
Best AKR1A1 / Aldo-keto reductase family 1 member A1 IHC Antibodies
A04855 has an IHC image from paraffin-embedded human tonsil carcinoma tissue (A04855 image caption). Its listed reactivity covers human, mouse, and rat (A04855 catalog); no IF image is supplied (A04855 catalog).
Real IHC data
Anti-AKR1A1 Antibody
Cat # A04855
A04855 is listed for IHC and reacts with human, mouse, and rat (A04855 catalog). Its IHC image shows paraffin-embedded human tonsil carcinoma tissue stained at 1:50, alongside a PBS-for-primary negative control (A04855 image caption).
Which to pick: For tissue IHC, choose A04855, a rabbit polyclonal antibody listed for IHC (A04855 catalog) with a paraffin-section image; the fixative is unreported (A04855 image caption). No SKU is listed for IF/ICC or has an IF image (A04855 catalog). For cross-species IHC, A04855 lists human, mouse, and rat reactivity (A04855 catalog), while its supplied IHC image documents human tissue only (A04855 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P14550 (AK1A1_HUMAN, Aldo-keto reductase family 1 member A1).
- Human Protein Atlas. AKR1A1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. AKR1A1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. AKR1A1 antibody validation summary (4 antibodies).
- Aldo-keto reductase family 1 member A1 (AKR1A1) exerts a protective function in alcohol-associated liver disease by reducing 4-HNE accumulation and p53 activation. Cell & bioscience 2024 — PMC10837880.
- A novel osteoporosis model with ascorbic acid deficiency in Akr1A1 gene knockout mice. Oncotarget 2017 — PMC5352327.
- Sexually Dimorphic Expression of eGFP Transgene in the Akr1A1 Locus of Mouse Liver Regulated by Sex Hormone-Related Epigenetic Remodeling. Scientific reports 2016 — PMC4834580.
- Metabolic reprogramming by the S-nitroso-CoA reductase system protects against kidney injury. Nature 2019 — PMC6318002.
- PubMed PMID:2498333 — UniProt-cited evidence.
- PubMed PMID:10393438 — UniProt-cited evidence.
- PubMed PMID:10486210 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AKR1A1 (Aldo-keto reductase family 1 member A1). Expected localisation Renal tubules show cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A04855 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil carcinoma tissue; fixative not specified (datasheet A04855); 2. Fixation: Image fixative and duration unreported (datasheet A04855); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-AKR1A1, 1:50-1:200 (datasheet A04855); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms annotated; processed chain starts at residue 2 (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt P14550, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A04855, PMC10837880, PMC5352327. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AKR1A1/immunohistochemistry-ihc-guide.