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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 tubules as a high-staining reference (HPA tissue IHC). This guide covers fixation consistency, antibody titration within 1:50–1:200 (datasheet), and interpretation of cytoplasmic and nuclear staining (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 | Cytoplasmic and nuclear staining in renal tubules (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear signal across renal tubule cells (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 fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | Regulation unspecified (UniProt) | |
| Isoform / epitope | No isoforms; chain 2–325; epitope location unreported (UniProt) |
Section 1
Recommended AKR1A1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with four published AKR1A1 paraffin-section protocols (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 antigen retrieval); use each article’s conditions when reproducing its method.
Section 2
What Is the Expected AKR1A1 Staining Pattern?
AKR1A1 is primarily cytosolic, with an apical cell membrane annotation and no transmembrane segment (UniProt P14550). In paraffin-section IHC, expect the clearest staining in kidney tubular cells, including cytoplasmic and nuclear staining; hepatocytes and several glandular or respiratory epithelial populations show weaker staining (HPA tissue IHC). HPA rates the tissue pattern Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Kidney tubules show strong cellular staining, with visible cytoplasm and nuclei. | This matches the High tubular-cell signal and cytoplasmic and nuclear profile reported for kidney (HPA tissue IHC). Compare neighboring cell populations separately: the HPA observation identifies tubular cells and does not establish that every cell in the section should stain equally (HPA tissue IHC). |
| Staining is exclusively extracellular or forms a sharp membrane-only outline, without convincing intracellular signal. | That distribution needs investigation: cytosol is the primary location (UniProt P14550; HPA subcellular ICC-IF), and kidney IHC includes cytoplasm and nuclei (HPA tissue IHC). An apical membrane annotation exists, so focal apical staining alone should not be dismissed; assess the full cellular pattern and controls before calling artefact (UniProt P14550). |
| The strongest signal appears in a cell population listed as undetected, while expected positive cells are weak. | For example, HPA reports no detectable staining in heart cardiomyocytes or cervical squamous epithelial cells (HPA tissue IHC). Unexpected signal raises cross-reactivity or detection-background concerns; it does not prove either cause. Recheck cell identity, morphology and controls alongside a kidney tubular-cell section (HPA tissue IHC; general IHC practice). |
| Brown color spreads across stroma, empty spaces or most cells without readable cellular boundaries. | Diffuse signal cannot establish AKR1A1 localization. Review a no-primary control for detection background and assess blocking, washes and chromogen development (general IHC practice). The intended comparison is cellular staining in kidney tubules, rather than uniform color across the section (HPA tissue IHC). |
| Kidney tubular cells show no convincing signal. | This conflicts with the High tubular-cell staining reported by HPA, but one negative section cannot identify the failed step (HPA tissue IHC). Check tissue preservation, antibody preparation, retrieval settings and detection controls using the IHC-validated antibody's instructions (general IHC practice). Target-specific fixation sensitivity is unreported in the supplied sources. |
💡Expected AKR1A1 appearanceCall a result positive when kidney tubular cells show High cytoplasmic and nuclear staining (HPA tissue IHC); diffuse acellular color or membrane-only outlining warrants a background or localization check (general IHC practice; UniProt P14550).
| Tissue and cell selection | Kidney tubular cells provide the strongest listed IHC reference (High); hepatocytes, bronchial respiratory epithelium and lung macrophages are Medium (HPA tissue IHC). HPA lists several undetected cell populations, so score the specified cells rather than declaring an entire organ positive or negative (HPA tissue IHC). |
| Compartment assignment | UniProt places AKR1A1 in the cytosol and at the apical cell membrane, with no transmembrane segment (UniProt P14550). HPA kidney IHC describes cytoplasmic and nuclear expression (HPA tissue IHC). These observations support checking intracellular signal while allowing nuclear and focal apical signal in the appropriate context. |
| Strength of tissue evidence | The tissue profile is Approved but has medium agreement between staining and RNA expression (HPA tissue IHC). Four listed antibodies have Approved IHC status (HPA antibodies). Treat this as support for the reported pattern, while retaining morphology and controls when interpreting an unexpected distribution (general IHC practice). |
| IF/ICC Q&A: Where should cellular fluorescence appear? | Mainly in cytosol, with additional nucleoplasmic signal (HPA subcellular ICC-IF). HPA lists A-431, U-251MG and U2OS ICC-IF images (HPA subcellular ICC-IF). This is localization context for the separate IF/ICC guide; the paraffin-section tissue pattern here comes from HPA tissue IHC. |
| Retrieval and detection checks | Use the IHC-validated antibody's documented paraffin-section workflow as the starting point (general IHC practice). Retrieval, blocking and detection settings can be checked against controls when a run fails (general IHC practice). The supplied UniProt and HPA records do not identify an AKR1A1-specific retrieval requirement or fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubular cells are negative across the section. | A failed staining run is possible; HPA reports High tubular-cell staining (HPA tissue IHC). | Check the IHC-validated antibody's instructions, reagent preparation, retrieval and detection controls (general IHC practice). Do not infer AKR1A1-specific fixation sensitivity from this result. |
| Nuclei stain along with tubular cytoplasm. | This can fit the reported kidney pattern, which includes nuclear expression (HPA tissue IHC). | Assess whether staining is confined to plausible cells and whether background controls are clean before treating nuclear signal as nonspecific (HPA tissue IHC; general IHC practice). |
| Color appears in no-primary controls or across acellular areas. | Detection background can produce color without target recognition (general IHC practice). | Review endogenous enzyme blocking where relevant to the chromogen, secondary detection, washes and development time (general IHC practice). Reassess AKR1A1 only after cellular boundaries become interpretable. |
| Only a continuous cell-surface rim is visible. | A membrane-only pattern does not reflect the mainly cytosolic localization, although UniProt also notes an apical membrane location (UniProt P14550; HPA subcellular ICC-IF). | Inspect the apical orientation and intracellular staining, then compare with kidney tubules and controls before assigning a compartment (HPA tissue IHC; general IHC practice). |
| A listed undetected cell type stains strongly. | The result conflicts with that specific HPA observation and could reflect cross-reactivity or detection background; its cause remains unproven (HPA tissue IHC; general IHC practice). | Confirm the cell type, examine a no-primary control and compare an expected positive population from the same run (HPA tissue IHC; general IHC practice). |
| A medium-level tissue looks faint beside kidney. | That difference may match HPA levels: kidney tubular cells are High, while hepatocytes and bronchial respiratory epithelial cells are Medium (HPA tissue IHC). | Score the named cell populations and compare sections processed together before changing staining conditions (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: cells in tubules should stain (HPA: High in kidney cells in tubules). Use esophagus squamous epithelial cells as a negative tissue (HPA: Not detected in esophagus squamous epithelial cells); on the kidney slide, assess unstained adjacent cells as background without assuming a particular cell type is AKR1A1-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) slide, a matched rabbit IgG isotype control, and an AKR1A1-knockout biological negative (selected IHC caption: rabbit primary detected with goat anti-rabbit IgG). In kidney, check endogenous peroxidase and biotin background when using the caption’s avidin-peroxidase detection method (selected IHC caption: avidin-peroxidase; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A04855 paraffin-section caption also does not report a fixative. Optimize antigen retrieval for paraffin IHC empirically (standard IHC practice). Frozen-section performance is unreported, while ICC-IF images are available for A-431, U-251MG and U2OS (HPA: subcellular); kidney tubule staining warrants particular attention to endogenous biotin background with avidin-based detection (HPA: High in kidney cells in tubules; selected IHC caption: avidin-peroxidase; standard IHC practice).
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 by checking retrieval, controls, cellular localisation and scoring before drawing biological conclusions.
How should I retrieve AKR1A1 in paraffin sections when staining is weak?
Use citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min for paraffin-section AKR1A1 IHC (page antigen-retrieval setting). Run a kidney section alongside the test section because tubular cells show high AKR1A1 staining (HPA: High in kidney tubular cells). Keep section thickness, heating, cooling and chromogen development consistent across slides when comparing staining (standard IHC practice). If the kidney control stains but the test section does not, check tissue preservation and cell composition before intensifying retrieval (standard IHC practice; HPA: kidney tubular-cell expression). Document any retrieval change as a separate condition because it can alter both signal and background (standard IHC practice).
Could fixation explain weak or variable AKR1A1 staining?
AKR1A1-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section image does not state a fixative (A04855 caption: fixative not stated). Record the fixative and fixation duration for each specimen, then compare sections processed together under the same citrate pH 6.0 retrieval condition (standard IHC practice; page antigen-retrieval setting). Uneven or prolonged fixation can change antigen accessibility in paraffin sections, but that is general IHC guidance rather than an established AKR1A1 effect (standard IHC practice). Include a similarly processed kidney control to help distinguish a failed staining run from a specimen-dependent result (HPA: High in kidney tubular cells; standard IHC practice).
Should AKR1A1 staining appear in the cytoplasm, nucleus or apical surface?
Expect a substantial cytoplasmic component because AKR1A1 is annotated in the cytosol, while an apical membrane location is also listed (UniProt P14550: subcellular location). Nuclear staining cannot automatically be rejected: tissue IHC reports cytoplasmic and nuclear expression in renal tubules, and subcellular IF reports additional nucleoplasmic localisation (HPA: renal-tubule profile; HPA: subcellular localisation). The selected paraffin-section image likewise describes cytoplasmic and nuclear staining at 1:50 (A04855 caption). Score compartments separately, comparing each with the same-slide primary-omission control and tissue morphology (A04855 caption: PBS primary-omission control; standard IHC practice). Diffuse chromogen outside intact cells should be assessed as possible background rather than assigned a subcellular location (standard IHC practice).
How can epitope uncertainty affect AKR1A1 staining in paraffin sections?
The supplied record lists 0 isoforms and an AKR1A1 chain spanning residues 2–325, but it does not map the catalog antibody’s epitope (UniProt P14550: isoforms and processing; supplied antibody evidence). Therefore, a compartment difference between specimens cannot be attributed to isoform-specific recognition from these data (UniProt P14550: isoforms; standard interpretation practice). AKR1A1 has annotated acetylation, phosphorylation and lysine modifications; their effect on this antibody’s paraffin-section binding is unknown without an epitope map (UniProt P14550: modified residues; supplied antibody evidence). Compare staining with a primary-omission control and a kidney control before assigning biological meaning to an unexpected pattern (A04855 caption; HPA: High in kidney tubular cells; standard IHC practice).
How should I assess an AKR1A1 immunofluorescence result alongside this IHC guide?
Treat IF/ICC as a separate application when comparing it with chromogenic paraffin-section IHC; the selected tissue image documents IHC staining at 1:50, not an IF protocol (A04855 caption). For IF multiplexing, pair AKR1A1 with a marker identifying the expected cell population, such as a renal tubular marker in kidney, and check each single-marker control (HPA: High in kidney tubular cells; standard IF practice). Choose a fluorophore channel after inspecting unstained tissue autofluorescence and avoid a channel with strong native signal (standard IF practice). Use permeabilisation when assessing the cytosolic or nucleoplasmic signal, and evaluate any apical signal against membrane morphology because AKR1A1 has no transmembrane segment (HPA: subcellular localisation; UniProt P14550: topology and subcellular location; standard IF practice).
What should I check when AKR1A1 chromogenic staining is widespread or patchy?
AKR1A1 is widely expressed, so widespread cellular staining alone does not establish a failed assay (UniProt P14550: tissue specificity). Compare the specimen with a primary-omission section; the selected paraffin-section image used PBS instead of primary antibody for its negative control (A04855 caption). If the omission control develops chromogen, review the 3% hydrogen-peroxide block for 10 min, blocking, washes and detection reagents as general peroxidase-IHC workflow variables (standard IHC practice). Examine whether precipitate, tissue folds, section edges or necrotic areas account for patchy colour before changing antibody conditions (standard IHC practice). Reassess cellular staining against a kidney tubular-cell control processed in the same run (HPA: High in kidney tubular cells; standard IHC practice).
How should I quantify AKR1A1 staining across paraffin-section specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because AKR1A1 staining can be cytoplasmic and nuclear in renal tubules (HPA: renal-tubule profile). For a cell-based comparison, record the percentage of positive target cells and intensity on a 0–3 scale, then calculate an H-score from 0–300 (standard IHC practice). Normalise counts to the number of eligible, intact cells in the same annotated compartment; use positive-cell density per mm² only when sampled tissue area is comparable (standard IHC practice). Apply one threshold, sampling rule and chromogen-development condition across specimens, excluding folds and necrosis by predefined criteria (standard IHC practice). Report cytoplasmic and nuclear scores separately rather than merging unlike patterns into one value (HPA: renal-tubule profile; standard IHC practice).
How can I distinguish true AKR1A1 staining from artefact?
Give greatest weight to staining within intact cells in an expected population, such as kidney tubular cells, which show high AKR1A1 expression (HPA: High in kidney tubular cells; standard IHC practice). Cytoplasmic staining fits the supported cytosolic location, while nuclear signal needs contextual assessment because renal-tubule IHC and subcellular IF also report it (HPA: renal-tubule profile; HPA: subcellular localisation). Be cautious with staining restricted to cut edges, necrosis or cells outside the annotated population, especially when morphology is poor (standard IHC practice). Check a primary-omission section for nonspecific detection and endogenous peroxidase activity before interpreting chromogen as AKR1A1 (A04855 caption: PBS primary-omission control; standard IHC practice). The tissue-IHC profile has Approved reliability with medium staining–RNA consistency, so confirm unexpected patterns with an independent method (HPA: reliability description; standard IHC practice).
Boster reagents
Best AKR1A1 / Aldo-keto reductase family 1 member A1 IHC Antibodies
One AKR1A1 antibody has real IHC data from a human paraffin section (A04855 image caption); its listed reactivity also includes mouse and rat (catalog: A04855).
Real IHC data
Anti-AKR1A1 Antibody
Cat # A04855
A04855 will render with an IHC image of a human paraffin section at 1:50, showing cytoplasmic and nuclear staining (A04855 image caption). Its listed applications are IHC and WB, and its listed reactivity is human, mouse and rat (catalog: A04855).
Which to pick: Choose A04855 for tissue IHC: it is a rabbit antibody with a listed IHC dilution of 1:50–1:200 and a paraffin-section image (catalog: A04855; A04855 image caption). For mouse or rat IHC, A04855 has listed species reactivity, but the supplied image shows only human tissue (catalog: A04855; A04855 image caption). No IF/ICC application or image is supplied for A04855, and the paraffin-section caption does not report the fixative (catalog: A04855; 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 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). 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 fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms; chain 2–325 ; epitope location unreported (UniProt); Regulation: Regulation unspecified (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.