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AKR1B10 / Aldo-keto reductase family 1 member B10 · IHC design guide
Design Immunohistochemistry for AKR1B10
Plan AKR1B10 staining in paraffin sections using the cytoplasmic pattern reported in gastrointestinal and gallbladder tissue (HPA tissue IHC). This guide covers controls, staining interpretation, and the potential effect of secretion on tissue localisation (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AKR1B10 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); lysosomal secretion (UniProt) | |
| Staining pattern | Enterocytes and glandular cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02976) | |
| Positive control | Small intestine+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Secretion may separate protein staining from RNA location (HPA tissue IHC) | |
| Regulation | Regulation not specified (UniProt) | |
| Isoform / epitope | No isoforms annotated; no transmembrane epitope split (UniProt) |
Section 1
Recommended AKR1B10 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with 4 published AKR1B10 IHC protocols covering liver, endometrial, and esophageal tissues (PMC5011670; PMC6201704; PMC8305663; PMC11663324).
| Sample | Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02976) |
| Fixation | Image fixative and duration unreported (datasheet A02976); 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, 20 min (datasheet A02976) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02976) |
| Primary antibody | Rabbit anti-AKR1B10, 0.5-1μg/ml (datasheet A02976) |
| Primary incubation | Overnight at 4 °C (datasheet A02976) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02976) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AKR1B10-positive staining in enterocytes - Microvilli of small intestine (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in gastrointestinal tract and gall bladder. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A02976); three published protocols also use citrate pH 6 (PMC5011670; PMC6201704; PMC11663324).
Section 2
What Is the Expected AKR1B10 Staining Pattern?
AKR1B10 should show predominantly cytoplasmic staining in gastrointestinal epithelial cells, especially small intestinal enterocytes and stomach glandular cells (HPA tissue IHC: High). HPA rates the tissue profile Enhanced, while cautioning that the staining evidence includes antibodies targeting proteins from multiple genes (HPA tissue IHC). A membrane-spanning pattern is not expected from the protein sequence (UniProt O60218: no transmembrane segment); supported ICC-IF data also place AKR1B10 mainly in the cytosol (HPA subcellular).
| Strong cytoplasmic signal in small intestinal enterocytes or stomach glandular cells (HPA tissue IHC: High). | This fits the principal positive tissue pattern. Compare epithelial cells with neighboring structures on the same slide, and assess whether the signal is cell-associated rather than a uniform deposit (HPA tissue IHC; general IHC practice). |
| Predominantly nuclear signal, with little epithelial cytoplasmic staining (HPA tissue IHC: cytoplasmic profile). | Treat this as a localization mismatch requiring validation. HPA reports cytoplasmic tissue staining and supported cytosolic ICC-IF localization; UniProt also lists lysosomal localization and secretion, so unusual signal should be checked rather than assigned to AKR1B10 by appearance alone (HPA tissue IHC; HPA subcellular; UniProt O60218). |
| Strong staining in adipocytes or respiratory epithelial cells, while expected gastrointestinal epithelium is weak (HPA tissue IHC). | These cell types are listed as not detected in the HPA tissue examples. Consider cross-reactivity or endogenous detection activity, then compare a matched negative detection control and a known-positive section (HPA tissue IHC; general IHC practice). |
| Even haze across epithelium, stroma, and empty areas of the section (general IHC practice). | Diffuse background has little anatomical selectivity and cannot establish AKR1B10 expression. Review blocking, wash steps, primary antibody concentration, and detection controls before interpreting cell-specific signal (general IHC practice). |
| No epithelial signal in small intestine or stomach (HPA tissue IHC: High). | First question assay performance, because these are strong HPA positive examples. Check tissue preservation, retrieval and detection using the antibody's validated IHC conditions; a failed positive control makes a negative study section uninterpretable (HPA tissue IHC; general IHC practice). |
💡Expected AKR1B10 appearanceA convincing positive is high cytoplasmic signal in small intestinal enterocytes or stomach glandular cells (HPA tissue IHC), whereas dominant nuclear staining or uniform tissue-wide haze warrants investigation (HPA subcellular; general IHC practice).
| Cellular localization and secretion (UniProt O60218; HPA subcellular) | UniProt places AKR1B10 in lysosomes and describes lysosome-mediated secretion; HPA ICC-IF mainly supports cytosol, with additional plasma membrane localization. Interpret intracellular staining in the context of the tissue IHC cytoplasmic profile; secretion can complicate a direct RNA-to-protein comparison (UniProt O60218; HPA subcellular; HPA tissue IHC). |
| Tissue reference and antibody specificity (HPA tissue IHC; HPA antibodies) | HPA lists high staining in small intestine and stomach, medium staining in colon and gallbladder, and Enhanced overall tissue reliability. Its cross-gene antibody caution limits target assignment from a matching stain alone; antibody-level IHC statuses differ: CAB070163 Enhanced and HPA020280 Supported (HPA tissue IHC; HPA antibodies). |
| Adrenal discrepancy (UniProt O60218; HPA tissue IHC) | UniProt describes high adrenal expression, but HPA lists adrenal glandular cells as not detected by tissue IHC. Treat adrenal tissue as an uncertain staining reference and favor the HPA high-staining gastrointestinal examples for an IHC positive control (UniProt O60218; HPA tissue IHC). |
| IF/ICC Q: What pattern should an IF comparison show? (HPA subcellular) | A: Mainly cytosolic signal, with possible additional plasma membrane localization in the HPA ICC-IF record. The subcellular summary cautions that its antibodies target proteins from multiple genes; use this as localization context, not proof that an IHC signal is AKR1B10-specific (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive gastrointestinal epithelium is blank (HPA tissue IHC: High). | A failed staining workflow is possible; absence on this section alone does not establish absent protein (general IHC practice). | Review the IHC-validated antibody's stated retrieval and dilution, reagent activity, and detection controls; repeat with a known-positive section in the same run (general IHC practice). |
| Nuclear staining dominates (HPA tissue IHC: cytoplasmic profile). | The distribution conflicts with HPA tissue and supported cytosolic ICC-IF observations (HPA tissue IHC; HPA subcellular). | Compare a known-positive section and negative detection control; review antibody specificity before scoring nuclei as positive (general IHC practice; HPA tissue IHC cross-gene caution). |
| Broad staining appears in HPA not-detected cell types (HPA tissue IHC). | Cross-reactivity or endogenous detection activity may mimic a positive chromogenic signal (general IHC practice). | Check a negative detection control and compare cell types within the section; avoid assigning AKR1B10 from staining in an unexpected compartment alone (general IHC practice; HPA tissue IHC). |
| A diffuse chromogenic veil obscures epithelial boundaries (general IHC practice). | Nonspecific antibody binding, inadequate washes, or detection background are possible general IHC causes (general IHC practice). | Review blocking and washes, and compare the negative detection control before adjusting the antibody concentration within validated IHC conditions (general IHC practice). |
| Adrenal tissue is negative despite the UniProt expression summary (UniProt O60218). | UniProt reports high adrenal expression, whereas HPA records adrenal glandular cells as not detected by IHC (UniProt O60218; HPA tissue IHC). | Use HPA high-staining small intestine or stomach as the assay control; report the adrenal result with the source discrepancy rather than calling the run failed (HPA tissue IHC; UniProt O60218). |
| Membrane-edge or extracellular signal accompanies epithelial staining (HPA subcellular; UniProt O60218). | HPA ICC-IF reports additional plasma membrane localization, and UniProt describes secretion; these observations do not establish the identity of a tissue deposit (HPA subcellular; UniProt O60218). | Score the expected epithelial cytoplasmic pattern separately, check background controls, and avoid treating an isolated edge or deposit as definitive positivity (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing AKR1B10 staining
A02976 · IHCIHC analysis of AKR1B10 using anti-AKR1B10 antibody (A02976). AKR1B10 was detected in paraffin-embedded section of human intestinal cancer t
A02976 · IHCIHC analysis of AKR1B10 using anti-AKR1B10 antibody (A02976). AKR1B10 was detected in paraffin-embedded section of human liver cancer tissue
A02976 · IFIF analysis of AKR1B10 using anti-AKR1B10 antibody (A02976). AKR1B10 was detected in an immunocytochemical section of A549 cells. Enzyme ant
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/cerebral+cortex#imid_18963205
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/duodenum#imid_18963114
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/colon#imid_18963115
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/liver#imid_18748143
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA020280; green = AKR1B10, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA020280; green = AKR1B10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AKR1B10 IHC & IF
🧪Run small intestine first: enterocytes at the microvilli should stain (HPA: High in small-intestine enterocytes). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the positive slide, assess non-enterocyte cells for background rather than assuming they are confirmed AKR1B10-negative cells.
Positive control tissue: Small intestine (Enterocytes - Microvilli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AKR1B10 in A-549, Hep-G2, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a rabbit immunoglobulin isotype control matched to the primary antibody’s clonality, and AKR1B10 knockout material or a validated immunizing-peptide block as a biological negative (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and assess endogenous biotin in small-intestine sections because the reported detection uses a biotin–streptavidin system with DAB (caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02976 paraffin-section caption does not state a fixative (caption: fixative unreported). The reported IHC procedure uses citrate retrieval at pH 6 for 20 minutes, but this single procedure does not establish that retrieval is required under all conditions (caption: citrate retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; check small-intestine sections for endogenous-biotin background when using the reported biotin-based detection (caption: SABC/DAB; standard IHC practice).
HPA tissue IHC evidence for AKR1B10
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Small intestine | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Colon | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Duodenum | Enterocytes | 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 AKR1B10 IHC Tips
Troubleshoot AKR1B10 staining in paraffin sections by checking retrieval, tissue controls, cellular localisation and detection background before scoring (A02976 tissue-IHC caption; HPA tissue IHC).
Where should I start if AKR1B10 staining is weak in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes on paraffin sections (datasheet A02976; A02976 tissue-IHC caption). Compare a suspected weak section with a same-run small-intestine control, where enterocyte staining is reported as high (HPA tissue IHC). Keep heating, cooling and section thickness consistent between runs so changes in staining can be attributed to the retrieval trial (standard IHC practice). The product image documents staining of paraffin-embedded human intestinal cancer tissue after this retrieval, but its fixative is unreported (A02976 tissue-IHC caption). If staining remains weak, test an alternative retrieval condition on adjacent sections while retaining the documented condition as the reference (standard IHC practice).
Can I change fixation to recover a weak AKR1B10 IHC signal?
AKR1B10-specific sensitivity to fixation is unknown because the selected paraffin-section caption does not state a fixative (A02976 tissue-IHC caption). Record the fixative, fixation duration and processing history before attributing a weak result to the antigen or antibody (standard IHC practice). Compare adjacent sections processed together, then alter one processing variable at a time while holding citrate retrieval at pH 6 constant (datasheet A02976; standard IHC practice). Include a positive tissue control in each run; small-intestine enterocytes have reported high staining (HPA tissue IHC). Neither the reported tissue distribution nor the protein's topology establishes a fixation effect for this antibody (HPA tissue IHC; UniProt O60218 topology).
Is apical or extracellular staining plausible for AKR1B10?
Score cellular staining first: gastrointestinal and gallbladder expression is described as cytoplasmic in tissue sections (HPA tissue IHC). High staining is reported in small-intestine enterocytes at the microvilli, so annotate any apical pattern separately from diffuse cytoplasm (HPA tissue IHC). A supported cytosolic location and an additional plasma-membrane location appear in subcellular data, with a warning about antibody cross-reactivity (HPA subcellular). Lysosomal localisation and secretion through a lysosome-mediated pathway are also annotated, without a transmembrane segment (UniProt O60218). Require appropriate tissue controls and a coherent cell-associated pattern before treating isolated luminal staining as specific (standard IHC practice; HPA tissue IHC).
Could an isoform or epitope explain discordant AKR1B10 staining?
No protein isoforms are annotated for the 316-amino-acid AKR1B10 chain, so an isoform explanation lacks support here (UniProt O60218). The record lists no signal peptide, transmembrane segment or glycosylation sites, but does list acetyllysines at positions 125 and 263 (UniProt O60218). The antibody epitope is not supplied, so these features cannot establish whether fixation or retrieval masks its binding site (A02976 tissue-IHC caption; UniProt O60218). Treat disagreement between antibodies cautiously because the tissue and subcellular profiles warn that antibodies may recognise proteins from multiple genes (HPA tissue IHC; HPA subcellular). Compare staining patterns in matched positive and low-expression tissues before assigning the discrepancy to AKR1B10 biology (HPA tissue IHC; standard IHC practice).
How should I follow up an IHC pattern with AKR1B10 immunofluorescence?
Use IF as a separate follow-up: the selected product evidence documents chromogenic staining in paraffin sections, without an IF fixation or permeabilisation condition (A02976 tissue-IHC caption). Multiplex AKR1B10 with a marker identifying the epithelial cell population being evaluated, such as the reported enterocytes or glandular cells (HPA tissue IHC; standard IF practice). Choose a fluorophore channel after checking unstained tissue autofluorescence, and include single-stain controls for bleed-through (standard IF practice). If probing an intracellular epitope, compare permeabilised and nonpermeabilised preparations; the epitope is unspecified despite supported cytosolic localisation (HPA subcellular; standard IF practice). Interpret membrane or extracellular signal cautiously given the cross-reactivity warning and annotated secretion (HPA subcellular; UniProt O60218).
What should I check when brown signal appears outside expected AKR1B10-positive cells?
The selected image used 10% goat serum blocking, a biotinylated secondary antibody, streptavidin-biotin detection and DAB (A02976 tissue-IHC caption). Run a no-primary control to assess detection-system staining and inspect whether brown signal follows tissue structure rather than cells (standard IHC practice). With a biotin-based system, evaluate endogenous biotin; with DAB, check that endogenous peroxidase was blocked (A02976 tissue-IHC caption; standard IHC practice). Compare background against adipose-tissue adipocytes or bone-marrow hematopoietic cells, where staining was not detected in the reference profile (HPA tissue IHC). Reassess primary concentration and washing if diffuse background persists; the documented image used 1 μg/mL overnight at 4°C (A02976 tissue-IHC caption; standard IHC practice).
How should I quantify heterogeneous AKR1B10 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before analysis, since gastrointestinal staining is described as cytoplasmic and enterocyte microvilli can stain strongly (HPA tissue IHC). Record the percentage of positive cells and intensity grades, then calculate an H-score from 0–300 if graded intensity is reproducible (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable tissue and normalise cell counts to the relevant epithelial population (standard IHC practice). Exclude folds, damaged edges and necrosis using the same rules across sections (standard IHC practice). Report control-tissue results and batch conditions alongside scores so retrieval or detection differences are visible (HPA tissue IHC; datasheet A02976; standard IHC practice).
When is an apparent AKR1B10-positive area likely to be artefact?
A convincing result places staining in the expected cells and predominantly cytoplasmic compartment, as described for gastrointestinal tissue and gallbladder (HPA tissue IHC). Isolated nuclear staining, edge-only staining or brown deposits over necrosis need investigation before being scored as AKR1B10 (HPA tissue IHC; standard IHC practice). Compare the suspect area with a positive small-intestine control and a no-primary control in the same run (HPA tissue IHC; standard IHC practice). Check endogenous peroxidase and biotin effects because the documented image used DAB with a biotin-based detection system (A02976 tissue-IHC caption; standard IHC practice). Interpret unexpected membrane or extracellular signal cautiously because secretion is annotated and the subcellular antibody data carry a cross-reactivity warning (UniProt O60218; HPA subcellular).
Boster reagents
Best AKR1B10 / Aldo-keto reductase family 1 member B10 IHC Antibodies
A02976 has real IHC images from human paraffin-embedded intestinal and liver cancer tissue and an IF image from A549 cells (A02976 image captions).
Real IHC data
Anti-AKR1B10 Antibody ®
Cat # A02976
A02976 is listed for human IHC, IF and ICC (A02976 catalog: applications and reactivity). Its IHC images show paraffin-embedded intestinal and liver cancer tissue, and its IF image shows A549 cells (A02976 image captions).
Which to pick: For tissue IHC, choose A02976 for paraffin sections; its IHC captions document citrate pH 6 retrieval, but do not report the fixative (A02976 IHC image captions). For IF/ICC, choose A02976; both applications are listed, and its IF image shows A549 cells (A02976 catalog: applications; A02976 IF image caption). No cross-species choice is supported because A02976 lists Human reactivity only (A02976 catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O60218 (AK1BA_HUMAN, Aldo-keto reductase family 1 member B10).
- Human Protein Atlas. AKR1B10 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. AKR1B10 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the plasma membrane. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. AKR1B10 antibody validation summary (3 antibodies).
- Pretreatment AKR1B10 expression predicts the risk of hepatocellular carcinoma development after hepatitis C virus eradication. World journal of gastroenterology 2016 — PMC5011670.
- Immunohistochemistry Detects Increased Expression of Aldo-Keto Reductase Family 1 Member B10 (AKR1B10) in Early-Stage Hepatocellular Carcinoma. Medical science monitor : international medical journal of experimental and clinical research 2018 — PMC6201704.
- AKR1B1 and AKR1B10 as Prognostic Biomarkers of Endometrioid Endometrial Carcinomas. Cancers 2021 — PMC8305663.
- Aldo-keto reductase family 1 member B10 prevents esophageal squamous cell carcinoma from reactive carbonyl species-induced cell death and promotes its progression. Cancer cell international 2024 — PMC11663324.
- PubMed PMID:9565553 — UniProt-cited evidence.
- PubMed PMID:9765596 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AKR1B10 (Aldo-keto reductase family 1 member B10). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); lysosomal secretion (UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02976). Validated IHC antibody: A02976 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Small intestine (Enterocytes - Microvilli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02976); 2. Fixation: Image fixative and duration unreported (datasheet A02976); 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 , 20 min (datasheet A02976); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02976); 8. Primary antibody: Rabbit anti-AKR1B10, 0.5-1μg/ml (datasheet A02976); 9. Primary incubation: Overnight at 4 °C (datasheet A02976). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secretion may separate protein staining from RNA location (HPA tissue IHC); Isoform / epitope: No isoforms annotated; no transmembrane epitope split (UniProt); Regulation: Regulation not specified (UniProt). Evidence: UniProt O60218, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the plasma…, Datasheet A02976, PMC5011670, PMC6201704. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AKR1B10/immunohistochemistry-ihc-guide.